cng n thi Thc tp Ha sinh 1 (Nm 2011 2012)

CNG N THI THC TP HA SINH 1

NI DUNG THI THC TP: I. L THUYT THC TP: Trc nghim + Cu hi ngn - Nguyn tc; - C ch; - Vai tr thuc th; - Nhn din, bin lun, gii thch kt qu. 1. Phn ng tm Cetone trong nc tiu. 2. nh lng lipid trong huyt thanh (HT). 3. Phn ng Nynhydrin. 4. Phn ng Biuret. 5. Tm Protein trong nc tiu bng 2 phng php: ng kt + Heller. 6. Phn ng Molish. 7. Phn ng Fehling. 8. Phn ng Seliwanoff. 9. Phn ng thy phn Saccharose. 10.Tm Glucose trong NT bng PP. Benedict. 11.Xc nh hot Amylase trong NT. 12.Tm sc t mt trong NT. 13.Tm mu trong NT. 14.Tm mui mt trong NT. 15.Xt nghim cn lng NT. 16.S dng giy nhng NT 3 thng s. 17.Cc cng thc tnh Acid Uric, Ure, Creatinine mu & NT. 18.Nng bnh thng cc cht trong mu v NT. 19.Cch ly v bo qun mu mu v NT. II. THC HNH: Bt thm => Chy trm 1. Tm th Cetone trong NT. 2. Tm Protein trong NT (PP. Heller). 3. Tm mu trong NT. 4. Tm sc t mt trong NT. 5. Tm mui mt trong NT. 6. Tm Glucose trong NT (PP. Benedict).

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7. Xc nh hot Amylase trong NT (PP. Wolhgemuth). 8. S dng giy nhng NT 3 thng s. 9. Cn lng NT: Tm & xc nh mt s TP trong cn lng NT.

TM TT L THUYT:
1. P TM CC TH CETONE TRONG NC TIU: (Tr. 15) 1.1. Nguyn tc: Sodium nitroprussiate tc dng vi cc cht Cetone cho phc hp mu tm, phn ng ny xy ra trong mi trng kim. OHCetone + Natri nitroprussiate -------------------- Phc cht c mu tm Ch : Cc th Cetones bao gm: + Acid aceto acetic; + Acid -hydroxy butyric; + Acetone. 1.2. Thuc th: - Sodium nitroprussiate 10% trong nc; - Acid acetic kt tinh; - NH4OH m c. Ch : vai tr ca Acid acetic trong p ny l: gi cho phc cht c mu tm bn. Nu ko c Acid acetic th p cng xy ra nhng mu tm ca phc cht s mt i nhanh chng. 1.3. Nhn nh v bin lun: - Nhn nh: + Xut hin vng mu tm c th Cetones trong NT (+); + Ko xut hin vng mu tm Ko c th Cetones trong NT (-). - Bin lun: + Bnh thng: . Nng Cetones trong mu rt thp (# 1 mg/dL); . Khng c Cetones trong NT;

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. C th c Cetones trong NT khi nhn i, ch n ngho Glucid, giu Lipid. + Bnh l: c Cetones trong NT trong cc trng hp: . Bnh i tho ng nng hoc iu tr bng Insulin ko liu BN e da b hn m; . Nhn i lu, nn nhiu; . Vn ng c nhiu, HC Cushing, + Khi nng Cetones/mu > 70 mg% xut hin Cetones/ NT. + Khi nng Cetones/mu > 100 mg% hi th c mi Cetones. 2. NH LNG LIPID TRONG HUYT THANH (Tr. 14) (K thut Sulfo Phospho Vanilic): 2.1. Nguyn tc: Trong MT Acid Sulfuric v Acid Phosphoric, Lipid tc dng vi Vanilin cho phc cht mu hng. 2.2. Thuc th: - Acid sulfuric m c; - DD Cholesterol mu (200 mg%) pha trong Acid acetic; - Thuc th Vanilin trong H3PO4. 2.3. Nhn nh, bin lun, gii thch kt qu: - Cng thc:

+ D: mt quang ca ng th; + D0: mt quang ng chun; + 4 = 2 (g/L) x 2: nng Cholesterol chun (2 g/L) x t l pha long. - Bin lun: + Bnh thng: Lipid ton phn/HT: 4 7,5 g/L. + Th/ sinh l: tng sau ba n nhiu m, c thai, sau khi sinh, cho con b; Gim khi h/ th lc nhiu. + Th/ bnh l: . Tng Lipid mu nguyn pht: bnh di truyn (cha r nguyn nhn) c 5 loi

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. Tng Lipid mu th pht: tiu ng, vim gan, thn m, va x m, ph nim, 3. PHN NG NINHYDRIN: (Tr. 28 29) 3.1. Nguyn tc: DD Protein, Peptide hoc Acid amin khi un nng vi Ninhydrin 0,2% s cho phc cht mu xanh tm. 3.2. C ch:

Ninhydrin l mt cht oxy ha nn c th to nn phn ng Carboxyl Oxy ha ca Acid amin vi nc, cui cng cho ra CO2, NH3, mt Aldehyde ngn i mt C so vi gc Acid amin ban u v Ninhydrin b kh. Sau , Ninhydrin b kh li tip tc tc dng vi NH3 va c phng thch v kt hp vi mt phn t Ninhydrin th hai to thnh Sp ngng kt c mu xanh tm. 3.3. Bin lun kt qu:

Vn tc phn ng, cng mu xanh tm ph thuc vo s lng nhm COOH v -NH2 t do (Acid amin t do > DD Peptide > DD lng trng trng). 3.4. ngha ca phn ng:

y l phn ng chung cho cc Protid v Acid Amin t do. P ny cho php nhn dng tt c cc AA c nhm NH2 v COOH t do. Ngoi tr Prolin v OH Prolin tc dng vi Ninhydrin cho mu vng. 4. PHN NG BIURET: (Tr. 30 31) 4.1. Nguyn tc: Protein tc dng vi Cu++ trong MT kim to phc cht c mu tm hng, p xy ra l do cc LK Peptide. Ti sao gi l p Biuret? S d gi l P Biuret l do cht Biuret (c nhm CO-NH, ging nh mt lk Peptide) cng cho p tng t, to phc hp c mu ging nh Protein. 4.2. ng dng v ngha ca P: - ng dng: nh lng Protein.

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- ngha: xc nhn cc lk Peptide. 5. TM PROTEIN TRONG NC TIU BNG 2 PP: NG KT & HELLER. (Tr. 32) 5.1. PP. ng Kt: (ta Protein bng Acid yu + un nng) Protein ha tan trong nc to thnh dd keo, trong , cc tiu phn Protein s tch in cng du v c lp o nc (hydrate ha). Nh tch in cng du nn cc tiu phn Protein y nhau v nh c lp o nc nn chng ngn cch vi nhau. V vy, dd keo bn vng. Nu lm mt 2 yu t trn th cc tiu phn Protein do chuyn ng s gp nhau, dnh vo nhau v to thnh nhng ht to ri kt ta. Trong PP. ng kt, to kt ta protein bng cch: - Dng Acid yu: a pH ca mi trng cha Protein v pHi ng in ca Protein lm mt lp in tch ca Protein. - un nng: lm mt lp o nc ca Protein. 5.2. PP. Heller (ta Protein bng acid mnh + khng un) Nguyn tc: Cc Acid v c mnh (HNO3, H2SO4, HCl, ) v cc acid hu c (Acid Trichloracetic, Acid Sulfosalicylic) c tc dng lm bin tnh v kt ta i a s Protein. Kt qu: to kt ta mu vng gia 2 mt phn cch. 6. PHN NG MOLISH: (Tr. 17) 6.1. Nguyn tc: Cc loi Glucid u cho phc cht MU TM vi dd Naphtol trong Acid sulfuric m c. 6.2. Thuc th Molish: DD Naphtol 1% trong cn 900. 6.3. Kt qu: - Cng mu ca cc ng: Fructose > Arabinose > Glucose > H tinh bt. - Do: Fructose v Arabinose c vng 5 cnh, trong mt Acid Sulfuric d b v vng to Furfural => kt hp Naphtol => phc cht Mu tm. Cn Glucose c vng 6 cnh bn hn; H tinh bt: kh bn
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6.4. ngha: - Phn bit glucid vi cc cht khc. - Phn bit c cc loi Glucid vi nhau. 7. PHN NG FEHLING: (Tr. 17 18) 7.1. Nguyn tc: Trong mt kim mnh, cc MS dng enediols (cht kh) khng bn, d dng kh cc kim loi nng nh Cu++, Ag+, Hg++. Cc ni i b ct t to thnh hn hp ng-acid. Cu++ MS + Base mnh enediols -----------------> Cu+ + HH ng-Acid (cht kh) OHCuOH to Cu2O (ta gch) 7.2. Kt qu: - Glucose, Fructose, Lactose c cha nhm OH bn Acetal c tnh kh to ta gch. - Saccharose v h tinh bt: ko cha OH bn Acetal ko c tnh kh ko to ta gch. 7.3. ng dng, ngha: - ng dng: tm ng trong NT. - ngha: khi P Fehling (+) kt lun: c ng/ NT v ng c tnh kh. 8. PHN NG SELIWANOFF: (c hiu cho Cetose) (Tr. 18 19) 8.1. Nguyn tc: Fructose v nhng Cetohexose khc to thnh Hydroxymethyl-furfural khi un nng vi Acid v c, cht ny tc dng vi Resorcinol cho phc mu .

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Cc Aldose cng c th to thnh Hydroxy-furfural khi un nng vi Acid v c, nhng phn ng xy ra chm. Do , P SELIWANOFF c tnh c hiu cho Cetose. 8.2. Thuc th Seliwanoff:

Gm: 0,05 gram Resorcinol (hay Resorcin) v 100 ml HCl pha long 1/3. 9. PHN NG THY PHN SACCHAROSE: (Tr. 19 20) 9.1. Nguyn tc: Saccharose khng c tnh kh, nhng khi thy phn bng acid th Saccharose bin thnh Glucose v Fructose u c tnh kh. 9.2. Kt qu: - ng cha dch thy phn ca Saccharose: P Fehling (+) tc c ta gch do G & F u c tnh kh; P Seliwanoff (+) tc cho phc cht mu do sp thy phn c Fructose. - ng cha dd Saccharose 1%: P Fehling (-) do Saccharose ko c nhm OH bn Acetal nn ko c tnh kh; P Seliwanoff (+) l do trong TT Seliwanoff c cha HCl pha long 1/3, trong K un cch thy 10 pht => Saccharose b thy phn => to Sp c Fructose. 10.TM GLUCOSE TRONG NC TIU (PP. BENEDICT): (bn nh lng) (Tr. 22 23) 10.1. Nguyn tc: Glucose c nhm Aldehyde s kh Cu++ thnh Cu+, to Oxyde ng (I) ta gch. 10.2. Thuc th Benedict: - c to thnh t 2 dd A & B; - Yu cu: phi ko c ta v bo qun c lu di. 10.3. Kt qu: - P (-): ch c mu xanh dng ca thuc th. - P (+): c mu xanh, kt ta t < 5 g/L. - P (++): un si 1 pht c kt ta gch nhiu hn, t mu xanh # 5 10 g/L.
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- P (+++): un si, ta gch nhiu # 10 20 g/L. - P (++++): ta mu nu sm ngay lc mi bt u un si > 20 g/L. 10.4. Phn bit: - P Benedict l p bn nh lng v: thng qua phn ng nh tnh, ngi ta c th c chng c lng Glucose c trong NT. - Phn bit TT Benedict & TT Fehling: TT Fehling ch c th nh tnh Glucose trong NT. 11.XC NH HOT AMYLASE TRONG NC TIU (PP. WOHLGEMUTH): (Tr. 63) 11.1. Nguyn tc: Dng phng php pha long dn NT tm lng enzyme ti thiu phn hy ht 2ml dd h tinh bt 1%0 370C trong 30 pht. Kim sot phn hy ca h tinh bt bng Iode. Tnh hot ca amylase theo n vi Wohlgemuth. nh ngha 1 n v Wohlgemuth: l lng Amylase ti thiu c kh nng thy phn 1ml dd h tinh bt 1%0 370C trong 30 pht. 11.2. Kt qu: - Chn ng biu din KQ: chn ng c mu trung gian hay khng mu ngay trc ng c mu xanh dng trc tin hin ra. - Cch tnh hot Amylase trong NT: Hot Amylase trong NT theo /v Wohlgemuth = pha long NT ca ng biu din KQ x 2 - Th d: ng 5 l ng xut hin mu xanh dng u tin chn ng 4 lm ng biu din KQ; ng 4 c pha long NT l 1/16 mL hot Amylase/NT = 16 x 2 = 32 /v Wohlgemuth. 11.3. Bin lun: - Ch : mu sc cc ng: + Cc ng no Amylase thy phn hon ton tinh bt thnh Maltose & Glucose: khng mu; + Cc ng no Amylase thy phn ko hon ton tinh bt to sp dang d Dextrin: mu trung gian;

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+ Cc ng no cn tinh bt: xanh dng. - Bnh thng: hot Amylase/NT: 16 32 /v Wohlgemuth (nhng c th thay i t 8 64 /v Wohlgemuth). Hot Amylase/NT cng phn nh hot Amylase/HT. - Bnh l: trong vim ty cp, Amylase niu > 200 /v (i khi ti 4000 5000 /v), t ti a vo lc 12 24h sau khi xut hin triu chng v tr v bnh thng sau t 2 3 ngy. 12.TM SC T MT TRONG NC TIU (K THUT FOUCHET): (trang 47) Sc t mt c ngha rng ch nhng sc t ca qu trnh thoi ha Hemoglobine. Sc t mt/NT l Bilirubine trong NT (Bilirubine trc tip). 12.1. Nguyn tc: Dng BaCl2 kt ta Bilirubine di dng mui khng tan Bari Bilirubinate. BaCl2 + Bilirubine ---------------> Bari bilirubinate (ko tan) Oxy ha mui ny bng FeCl3, bin Bilirubine thnh Biliverdine c mu xanh ve. 12.2. Thuc th Fouchet: gm c: - Acid Trichloracetic 25% 100mL. - DD FeCl3 10% 10 mL. 12.3. Bin lun KQ: - Bnh thng: ko c Bilirubine trong NT. - Bnh l: nu c Bilirubine/NT th l loi Blirubine trc tip. Gp trong cc bnh l gy vng da ti gan v sau gan. VD: vim gan siu vi, vim gan do nhim c ha cht, vng do tc mt. 13.TM MU TRONG NC TIU (PHN NG MAYER): (Tr. 48) 13.1. Nguyn tc:

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Hemoglobine (ngay c khi cha b bin tnh) c tc dng nh mt Peroxydase tc c kh nng gii phng Oxy hot ng t Hydroperoxyde (nc Oxy gi). Oxy ny c kh nng Oxy ha mt s thuc th cho mu c bit (Vd: Phenolphtalein b kh, Pyramidine, Pyridine, ). i vi Phenolphtalein, Oxy ny s Oxy ha Phenolphtalein dng kh to phc mu . 13.2. Thuc th Mayer: tp c cha Phenolphtalein 13.3. c kt qu, Bin lun: - P Mayer (+): KL c mu hoc Hb trong nc tiu (Nu mun bit chnh xc phi soi nc tiu). - Soi NT: + Thy HC c MU trong NT; + Ko thy HC m Mayer (+) C Hb trong NT. - Nhn i th: + NT c mu: mu ; + NT c Hb: mu x x. - Bnh thng: ko c mu hay Hb trong NT. - Bnh l: + Tiu ra Hb trong thiu G6PD, truyn nhm nhm mu, + Tiu ra mu: sn ng tiu, lao ng tiu, ung th bng i, ung th thn. 14.TM MUI MT TRONG NC TIU (PHN NG HAY): (Tr. 48) 14.1. Nguyn tc: Cc mui kim ca Acid mt lm gim r rt sc cng b mt ca NT. Dng lu hunh thng hoa pht hin hin tng ny. 14.2. Thuc th: bt lu hunh thng hoa 14.3. c kt qu: - P.. (+): lu hunh ri xung y ng nghim; - Nu c lng nh mui mt th sau 15 pht, lu hunh ch dn thnh lp mng m ko ri xung y ng nghim v phi g nh thnh ng lu hunh mi ri xung y ng nghim;

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- P.. (-): g nh m lu hunh ko ri xung y ng nghim. - Ch : p.. ny ko c hiu, n cho KQ (+) ngay c khi c Thymol, Phenol l nhng cht thng dng bo qun NT. 14.4. Bin lun: - Sc t mt, mui mt l 2 thnh phn quan trng ca mt, lin quan ch yu n bnh gan mt. - Vai tr ca mui mt: nh tng ha Lipid trong thc n => gip tiu ha cc cht m v cc Vitamin tan trong m. - Phn ng tm sc t mt, mui mt trong NT gip chn on cc bnh tc mt, vim gan: sc t mt, mui mt khng xung rut nn trn ra mu v ra NT. 15.XT NGHIM CN LNG NC TIU: (Tr. 54) 15.1. Nguyn tc: Trong NT c l lng cc thnh phn rt nh nh cc TB ca t chc, tinh th ha hc, Khi ly tm, cc tp ny tp trung li. Ly mt git cn NT ly tm em soi di knh hin vi gia lam knh v lammelle. 15.2. Kt qu: - Cch xem knh hin vi: + Vt knh 10X nhn tng qut quang trng; + Vt knh 40X xc nh loi t bo. - c kt qu: phn bit 2 loi cn: + Nhng cn hu c: l tt c nhng cht c cu to t bo v cc cht sng nh: hng cu, bch cu, TB biu m, trng KST, KST, + Nhng cn v c: l tt c nhng tinh th ca cc hp cht ha hc ko tan trong NT. - Nhng cn hu c: gm + Nhng yu t KT nh: hnh cu. VD: hng cu, bch cu, nm men, Trichomonas, tinh trng, + Nhng yu t KT trung bnh: t bo vi nhn. VD: TB biu m, TB bng quang, TB ca b thn, TB thn. + Nhng yu t c kch thc ln: tr niu (tr trong, tr ht, tr HC, tr BC, tr biu m, tr m)

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TR NIU: . Hnh dng: l nhng di hnh tr. . Di v chim ht b ngang ca vi trng vt knh 10X. . C th gp trong cc bnh ca ng thn, bn trong c cha y nhng HC, BC, TB hay cn ha hc. . C 3 loi KO GI L TR: tr gi; nhng si nhy; nhng cht bn khc: si bng, bong bng, ht tinh bt - Nhng cn v c: phn bit 3 loi: + Nhng cn VC bnh thng, kt tinh: Calcium Oxalate, Uric Acid, + Nhng cn VC bnh thng, v nh hnh: Amorphous phosphate (ht nh mu trng, thng gp trong NT kim), Urate v nh hnh (ht nh mu vng, vn li thnh tng m dy c, thng gp trong NT Acid). + Nhng cn VC ko bnh thng: tinh th Cystine, tinh th Tyrosin, Leucin - Cch biu th s lng HC, BC trong cn lng NT: + Bnh thng: HC < 10 / vi trng (40X) BC 2 3 / vi trng (40X) + Vi: 1 git cn (1/50 mL) Lammelle (20 x 20 mm) Vt knh 40X, th knh 5 hoc 6X 16.S DNG GIY NHNG NT 3 THNG S: (Tr. 72 74) 16.1. Cng thc: Dng loi giy th URITEST gm 3 thng s: pH Glucose Protein. 16.2. Ch nh: Bng th NT c sd chn on v theo di bnh tiu ng v cc bnh lin quan n thn, ng tit niu. 16.3. Cch dng: - Hng NT vo ng nghim (NT mi, cha quay ly tm); - Nhng ton b bng th vo ly ra nhanh ko nh u bng th trn thnh ng cho ht git NT tha. - c KQ:

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+ pH v Protein: c lin sau nhng; + Glucose c sau nhng 1 pht. Ch tn trng thi gian c KQ. 16.4. Xc nh KQ: - pH: ch th mu vng cam hi nu ty theo pH NT; bnh thng pH nc tiu 5 6. - Glucose: khi NT c glucose, vng ch th mu chuyn t mu TRNG NG hoc HI HNG (m tnh) TM (dng tnh). TM cng m => nng ng cng cao. Ch : + KQ yu or gi m khi: T trng NT cao; Nng Cetone cao; BN dng nhiu Vitamin C. + KQ gi dng khi: dng c ng NT c ln cht st trng: Javel, bt git, x bng. - Protein: c Protein: VNG SM (m tnh) XANH RU (dng tnh) Ch : KQ gi dng khi: + C Alcol trong NT (nhng mu ko bn); + Dng c cha NT c Acetate Amonium; + mt s BN sau khi dng QUININ. 16.5. Bin lun KQ: - pH: + Bnh thng: 5 6 (c th th/ 4 8). + pH thng thp nht lc sng sm, cao nht sau ba n. + pH < 4: Acid mnh. + pH > 8: kim. - Glucose: + Bnh thng: ko c Glucose trong NT. + Bnh l: tiu ng, Stress, vim tuy cp, HC Cushing, sau gy m, + c hiu: chuyn bit cho Glucose tr li KQ: c/ ko c Glucose. + nhy: 4 7 mmol/L Glucose (>= 0,5 g/L)/ - Protein: + Bnh thng: NT cha mt lng nh Albumin v globulin, khng gy (+). + Quan trng: NT phi c c c mt cch y .
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+ nhy: 0,15 0,30 g/L (Gram Albumin/Liter) (>= 0,15 g/L). 17.CC CNG THC TNH ACID URIC, UR, CREATININE MU & NT: (Tr. 67 72) 17.1. nh lng Acid Uric: Cng thc: - Huyt thanh, huyt tng: C (mol/L) n = 476 C (mg/dL) n = 8 - Nc tiu: C (mol/L) n = 5236 C (mg/dL) n = 88 ( nhn sn t l pha) Ch : Nc tiu: pha long t l 1 NT + 10 Nc ct. Bin lun: - Bnh thng: + Trong mu: Nam: 200 420 mol/L (3,4 7,0 mg/dL). N: 140 340 mol/L (2,4 5,7 mg/dL). + Trong nc tiu: 250 750 mg/24h hay 1,5 4,5 mmol/24h. Ch : khi nng Hb > 100 mg/dL hoc Bilirubine > 20 mg/dL c th nh hng ti kt qu Acid Uric. - Bnh l: Acid Uric trong mu v NT tng + Nguyn pht: bnh Goutte (trong mu: thng tng r rt, nhng trong NT c th gp giai on gim). + Th pht: trong cc bnh v mu (a HC, BC cp hoc mn, u hch); trong iu tr tia X, phng x, cht chng chuyn ha; torng bnh v thn (vim thn mn, suy thn). 17.2. nh lng Ur: Cng thc: - Huyt thanh, huyt tng: C (mmol/L) n = 13,3

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C (mg/dL) n = 80 - Nc tiu: C (mmol/L) n = 13,3 x 101 = 1343,3 C (mg/dL) n = 80 x 101 = 8080 Ch : Nc tiu: pha long 1 mL NT vi 100 mL nc ct. Bin lun: Ur l sn phm thoi ha quan trng nht ca Protein. - Bnh thng: + Ur mu vo khong 10 50 mg/dL (1,7 8,3 mmol/L). + Ur NT khong 20 35 g/24h (333 583 mmol/24h). - Thay i sinh l: theo n ung, theo tui. - Thay i bnh l: Course TT/ Tr. 70 17.3. nh lng Creatinine: Xem Course TT/ Tr. 71 (son ht kp oy.) 18.NNG BNH THNG CA CC CHT TRONG MU, NT Xem Course TT/ Tr. 75 82 19.CCH LY V BO QUN MU MU, NT: Xem Course TT/ Tr. 7 - 9

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