New methylene blue ndi kupaka utoto ma reticulocyte: utoto wosiyana ndi njira yogwiritsa ntchito maselo amoyo

Wophunzira akukonzekera kuwerengera ma reticulocyte, akuitanitsa methylene blue, akuusakaniza ndi magazi, ndipo sakuona pafupifupi chilichonse. Mnzake akugwiritsa ntchito botolo lolembedwa new methylene blue ndipo akuona ulusi wabuluu wooneka bwino mkati mwa ena mwa maselo ofiira a magazi. Wophunzira woyambayo akuganiza kuti utoto wa m’botolo lachiwirilo ndi watsopano kwambiri kapena wokhuthala kwambiri. Palibe chimene chili choona pa ziwirizi. Muli molekyulu yosiyana.

Kumvetsa molakwika kumeneku ndiko kumayambitsa kusokonezeka kwakukulu pankhaniyi. Dzinalo limapereka chithunzi chakuti new methylene blue ndi methylene blue yopangidwa bwino kuposa kale. Koma kapangidwe kake ka mankhwala kakusonyeza zosiyana. New methylene blue N ndi utoto wina wa phenothiazine wokhala ndi nambala yake ya Colour Index, dongosolo lake la magulu olowa m’malo pa molekyulu, ndiponso ntchito yake yokhazikitsidwa pa kafukufuku wa magazi. Kusiyana kumeneko kukamveka bwino, mbali zina za njirayi zimakhala zosavuta kumvetsa: chifukwa chake maselo ayenera kukhala amoyo popakidwa utoto, zimene zimawerengedwa ngati reticulocyte, malo a matupi a Heinz, ndiponso nthawi imene brilliant cresyl blue imagwiritsidwa ntchito.

Yankho lachidule

New methylene blue N, chinthu chogwiritsidwa ntchito pa kafukufuku wa magazi, nthawi zambiri imadziwika kuti ndi Basic Blue 24 m’kaundula wa PubChem wa new methylene blue, C.I. 52030, CAS 1934-16-3, yokhala ndi fomula C18H22ClN3S. Methylene blue wamba ndi methylthioninium chloride, Basic Blue 9, m’kaundula wake wa PubChem, C.I. 52015, CAS 61-73-4, yokhala ndi fomula C16H18ClN3S. Pa kapangidwe kake, new methylene blue ili ndi magulu awiri a methyl pa mphete yake ndi magulu a ethylamino a m’mbali pamalo amene methylene blue ili ndi magulu a dimethylamino. Kusiyana kumeneko kumasintha mmene utotowo umagwirira ntchito. Mabuku a kafukufuku wa magazi amafotokoza kuti methylene blue wamba sapaka ma reticulocyte bwino kwambiri, pamene new methylene blue imapaka utoto wakuya komanso wofanana bwino. Kuyerekezera kumeneku kwafotokozedwa mwachidule mu nkhani ya ScienceDirect yokhudza new methylene blue.

Inde, new methylene blue ndi utoto wa supravital. Kupaka utoto mwa supravital kumatanthauza kupaka utoto maselo amoyo atachotsedwa m’thupi koma asanakhazikitsidwe kuti asinthe pang’ono, monga tafotokozera mu ndemanga ya NCBI yokhudza mfundo za kupaka utoto mwa supravital. New methylene blue ndi brilliant cresyl blue onse ali m’gulu limeneli, ndipo muyezo wa CLSI wa ma reticulocyte umatchula new methylene blue mwachindunji ngati utoto wa supravital. Nthawi zina ophunzira amaika njirayi m’gulu limodzi ndi kupaka utoto mwa Romanowsky pa zopaka zopyapyala zokhazikitsidwa kale. Kuganiza motero sikugwira ntchito pano. Mukakhazikitsa maselo kaye, njirayi sigwiranso ntchito.

Kwa owerenga amene akutsatira mayina a mitundu ya utoto yofanana, kusiyana kwa methyl blue ndi methylene blue kumafotokoza kusokonezeka kwina kwakukulu pakati pa utoto wa triarylmethane wokhala ndi chaji yosavomereza ndi gulu la phenothiazine lokhala ndi chaji yovomereza. Chidule cha kupaka utoto m’labotale chimaika ntchito ya supravital pakati pa njira zogwiritsa ntchito zopaka zopyapyala zokhazikitsidwa kale, ndipo kalozera wa mawonekedwe a maselo a magazi amasonyeza pamene kuunika ma reticulocyte kumalowa pa kusiyanitsa mitundu ya maselo ambiri.

Chenjezo limodzi pa makatalogu: pali mtundu wina wa new methylene blue wokhala ngati mchere wapawiri wa zinc chloride, pansi pa CAS 6586-05-6. Choncho gawo lofotokoza njira liyenera kutchula mtundu wa mchere ndi wogulitsa, osati dzina lodziwika lokha.

Zimene kupaka utoto mwa supravital kumachita kwenikweni

Reticulocyte ndi selo lofiira la magazi lachinyamata limene lataya phata lake koma likadali ndi ribosomal RNA yotsala. Pa chitsanzo chonyowa chosapakidwa utoto, RNA imeneyo sioneka. Utoto wa supravital umalowa m’selo lamoyo ndipo umapangitsa RNA imeneyo kugwa kuchokera m’madzimadzi kapena kusonkhana kukhala tinthu tabuluu ndi ulusi. Zinthu zokhala ngati ukonde ndi ulusi zimenezi zimaoneka pogwiritsa ntchito mafuta pakati pa chitsanzo ndi lenzi ya maikulosikopu. Choncho ukondewo ndi chinthu chopangidwa ndi kupaka utoto, osati chigoba chomwe chinalipo kale chikungoyembekezera mtundu. N’chifukwa chake zinthu zosinthika m’ndondomekoyi, monga kuchuluka kwa utoto m’madzimadzi, chosunga pH, chiŵerengero cha magazi ndi utoto, kutentha ndi nthawi, n’zofunika mofanana ndi dzina la utotowo.

Pa ntchito yeniyeni, zimenezi zimafuna dongosolo lokhazikika: choyamba sakanizani magazi oikidwa choletsa kuundana ndi utoto, siyani kwa nthawi yake pa kutentha koyenera, kenako pangani zopaka zopyapyala ndi kuziona popanda kukhazikitsa maselo kapena kuwonjezera utoto wina wosiyanitsa. Kukhazikitsa maselo musanawapake utoto, kapena kuwonjezera utoto wosiyanitsa umene umabisa ulusi wosaoneka kwambiri, kumawononga uthenga umene kuwerengerako kumadalira.

Ndondomeko yogwira ntchito yowerengera ma reticulocyte ndi new methylene blue

Palibe kusakaniza kumodzi kogwiritsidwa ntchito kulikonse, choncho nenani kumene mwagwiritsa ntchito. Njira zitatu zolembedwa zimasonyeza kusiyanasiyana kwake.

Njira yakale ya mtundu wa Brecher imagwiritsa ntchito new methylene blue pafupifupi 0.5 percent yokhala ndi potassium oxalate. Imasakanizidwa ndi magazi a EDTA osakanizidwa bwino, pogwiritsa ntchito milingo yofanana, n’kusiyidwa kwa mphindi pafupifupi 10 mpaka 15 pa kutentha kwa m’chipinda. Pambuyo posakanizanso mofatsa, amakonza zopaka zopyapyala zooneka ngati mphero, kuziumitsa ndi mpweya, ndi kuziona popanda kukhazikitsa maselo pogwiritsa ntchito lenzi ya 100x ya mafuta. Ndondomeko ya NCBI yowerengera ma reticulocyte imafotokoza kayendedwe kameneka ndi chimango chowerengera chomwe chimagwiritsidwa ntchito.

Njira ya Dacie ndi Lewis imagwiritsa ntchito new methylene blue ya 1.0 percent m’madzi amchere okhala ndi citrate, yosakanizidwa ndi magazi ochuluka kuwirikiza 2 mpaka 4, n’kusungidwa pa 37 C kwa mphindi 15 mpaka 20 musanapange zopaka zopyapyala. Kukhazikitsa kwaposachedwapa kwa muyezo wofanana wa ndondomeko ya CLSI H44 kunagwiritsa ntchito madzi osakaniza osakhuthala kwambiri a 0.1 percent m’madzi amchere okhala ndi phosphate yosunga pH pa 7.4. Kusakanizako kunasiyidwa pa 20 C kwa mphindi 10 popanda kukhazikitsa maselo, monga tafotokozera mu kafukufuku wa 2026 wokhazikitsa muyezo wofanana wa NMB.

Malamulo owerengera ndi ofanana pa njira zonsezi. Werengani selo ngati reticulocyte ngati lili ndi tinthu tapadera topakidwa utoto wabuluu kapena zigawo za ulusi zosachepera ziwiri. Zimenezi zimachotsa tinthu tokha ta utoto womwe wangotsalira. Werengani maselo ofiira a magazi osachepera 1,000 pa ntchito yanthawi zonse ndipo perekani ma reticulocyte ngati chiŵerengero cha pa zana. Chiŵerengero cha pa zana chikakhala chochepa, ziwerengero zimafuna maselo ambiri. Ndemanga ya njira ya CLSI H44 imanena kuti ma reticulocyte akakhala pafupi ndi 1 percent, pamafunika maselo pafupifupi 4,400 kuti kusiyanasiyana kwa kuwerengera kufike pa 5 percent, mu ndemanga ya CLSI H44 ku De Gruyter. Disiki ya Miller yoyikidwa m’lenzi yoyang’aniramo imathandiza kuwerengera maselo ambiri mosavuta. Sefani utoto kapena konzani watsopano, chifukwa utoto umene wagwa kuchokera m’madzimadzi umafanana ndi tinthu tating’ono tomwe timakhala m’maselo.

Matupi a Heinz m’chitsanzo chomwecho

Chopaka chopyapyala chomwecho cha supravital chingasonyezenso matupi a Heinz. Amenewa ndi magulu a hemoglobin yomwe yasintha chifukwa cha oxidation ndi kutaya kapangidwe kake, osati RNA yotsala. Pa zitsanzo zopakidwa ndi new methylene blue, amaoneka ngati matupi ozungulira, nthawi zambiri limodzi m’selo ndipo pafupi ndi nembanemba, opakidwa utoto wabuluu wopepuka kuposa zinthu zokhala ngati ukonde ndi ulusi. Sasungika bwino maselo akakhazikitsidwa ndi kuwonjezedwa utoto wosiyanitsa, choncho yang’anani chopaka chopyapyala chosakhazikitsidwa. Ndondomeko yaposachedwapa yogwiritsa ntchito magazi a EDTA a 20 microliters ndi new methylene blue ya 1 percent yokwana 20 microliters, n’kusiya kusakanizako kwa mphindi 15, inawerengera maselo okhala ndi matupi a Heinz pakati pa maselo ofiira a magazi 250, mu kafukufuku wa PMC wofotokoza ndondomeko imeneyo.

Kusiyanitsa zinthu ziwiri kumathandiza kupewa kuwerenga molakwika. Choyamba, methyl violet ndi crystal violet zimapaka matupi a Heinz mwamphamvu kuposa new methylene blue, monga tafotokozera mu nkhani ya ScienceDirect yokhudza matupi a Heinz. New methylene blue ndi yosavuta kusankha pamene ma reticulocyte ndi matupi a Heinz akuunikidwa pamodzi, koma si utoto wamphamvu kwambiri wopangidwira kuunika matupi a Heinz okha. Chachiwiri, tinthu ta HbH tomwe timapezeka m’maselo a munthu wokhala ndi chibadwa cha alpha thalassemia timaoneka ngati madontho ang’onoang’ono ambiri m’selo lonse pambuyo pa kupaka utoto mwa supravital. Koma matupi a Heinz achizolowezi amakhala ochepa, akuluakulu, ndipo nthawi zambiri amakhala pafupi ndi nembanemba. Utoto wotsalira ndi chinthu chachitatu chofanana nawo: umakhala pa mulingo wina woyang’anirapo kusiyana ndi maselo ndipo umasiyanasiyana pakati pa malo oyang’anidwa.

Brilliant cresyl blue poyerekeza ndi new methylene blue

Mitundu yonseyi ndi utoto wa supravital wa ma reticulocyte, koma miyezo imakonda umodzi mwa iyo. Njira ya CLSI H44-A2 ya ma reticulocyte, yopangidwa pamodzi ndi International Council for Standardization in Haematology ndipo yotchulidwa pa tsamba la muyezo wa CLSI H44, imasankha kupaka magazi a EDTA ndi new methylene blue ngati njira yapamanja yoyerekezera. Maphunziro akale a kafukufuku wa magazi amavomerezana pa maonekedwe ake: new methylene blue imapaka ulusi utoto wakuya ndi wofanana bwino, pamene brilliant cresyl blue imasiyanasiyana kwambiri pakati pa magulu opangidwa ndi zitsanzo.

Udindo umenewo uyenera kufotokozedwa ndi malire ake. Malangizo a ICSH ounika zida zoyesera amavomereza kuunika ndi maikulosikopu pamanja pogwiritsa ntchito new methylene blue ngati njira yoyerekezera ndi muyezo, koma amati imadalira mmene woyang’anayo akuonera ndipo kulondola kwake n’kochepa poyerekeza ndi njira zina. Njira za muyezo zogwiritsa ntchito flow cytometry zimakondedwa ngati zilipo. Kuyerekezera kwa zitsanzo 100 mu 2022 kunapeza kuti brilliant cresyl blue inali ndi kupatuka kochepa pa zotsatira za chipangizo choyesera kuposa new methylene blue, monga tafotokozera mu kuyerekezera njira zopaka utoto kwa PMC. Chidule chovomerezeka kwa ophunzira ndi ichi: gwiritsani ntchito new methylene blue ngati njira yapamanja yoyerekezera yokhazikitsidwa ndi muyezo; tsimikizirani njira iliyonse ya brilliant cresyl blue yogwiritsidwa ntchito kwanuko poyiyerekezera ndi imeneyi kapena ndi njira ya makina; ndipo yembekezerani kusiyanasiyana kwakukulu kwa zotsatira za maikulosikopu yapamanja m’njira iliyonse. Mabuku akale okhazikitsa miyezo analimbikitsanso Azure B yoyeretsedwa ngati njira ina yokhala ndi mankhwala oyera kwambiri chifukwa magulu a utoto wogulitsidwa ankasiyanasiyana, mu kafukufuku wa Marshall wokhudza Azure B. Zimenezi zikufotokoza chifukwa chake mlingo wa kuyera kwa zinthu zoyesera umatchulidwabe pokambirana njira.

Kupewa zolakwa zofala

Zolakwa zinayi zimayambitsa kulephera kwa zitsanzo zambiri. Kuitanitsa methylene blue wamba m’malo mwa new methylene blue N kumapangitsa kupaka utoto kofooka chifukwa cha kapangidwe ka mankhwala, osati chifukwa cha njira yogwirira ntchito. Kukhazikitsa maselo kapena kuwonjezera utoto wosiyanitsa musanaone chitsanzo kumachotsa chizindikiro cha supravital. Kuwerengera utoto umene wagwa kuchokera m’madzimadzi ngati tinthu ta m’maselo kumakweza zotsatira; lamulo la tinthu tiwiri ndi utoto wosefedwa zimapewa zimenezi. Kuwerengera maselo ochepa kwambiri pamene chiŵerengero cha ma reticulocyte chili chochepa kumapereka nambala yooneka ngati yolondola kwambiri koma yokhala ndi malire otakata a chidaliro cha ziwerengero. Cholakwa chilichonse chimakhala ndi chizindikiro chofanana: wogwiritsa ntchito maikulosikopu amaimba mlandu maikulosikopuyo pa zimene zinatsimikizidwa kale pa gawo logwiritsa ntchito chinthu choyesera.