Chapter Blood

 RBC

 Platelet

 Hemostasis
 Section 1

Outline of the Blood

 Distribution of the Body Fluid

Intracellular fluid (ICF) 40%

Body fluid

Interstitial fluid 15%

Extracellular fluid Plasma 4%
(ECF) 20%
Other fluid 1%
(lymph, CSF)
Exchange between the capillary blood and the tissue flu
id is by formation and reabsorption of tissue fluid and b
y diffusion into and out of the capillary
 Functions of the Blood
 Transport
Nutrition and excretion, gases
 Maintain homeostasis of internal environme
nt
Ionic and osmotic balance, buffering
 Nervous-humoral regulation
 Protection
Hemostasis, anti-infection, immune reaction
 Heat balance
 Blood Volume
Blood volume= blood cells + plasma
7% to 8% of body weight.
Measurement of blood volume (formula)
Blood volume= plasma volume/(1-hemato
crit)
血浆 / （ 1- 红细胞比容）
 The total blood volume an average perso
n is approximately 5.5L
 Hematocrit
 Hematocrit
(the fractions of blood,
Packed Cell Volume,
PCV)
The percentage of the
original volume of blo
od then occupied by th
e erythrocytes is calle
 
d the hematocrit ( 比
容)
 Hematocrit
Male 40%—50%; female 37%—48%
Represent relative concentration of
RBC
Various with the distribution in vessels
Decrease in the patients being ill with
anemia
 Physics and Chemical
Properties of Blood

1. pH and Buffers
-pH 7.35 ~7.45 NaHCO3/H2CO3,
-buffer pairs in plasma
Na-prot./K-
prot.
KHb/HHb N
-buffer pairs in RBC aKHbO
2HPO 4/NaH22PO4
2/HHbO
K2HO4/KH2PO4
KHCO3/H2CO3
 Physics and Chemical
Properties of Blood
2. Viscosity （血液的粘度）
 Viscosity results from friction of liquid internal
molecule.
 water 1<plasma 1.6-2.4<blood 4-5
 Blood viscosity depend on hematocrit; plasma
viscosity depend on protein content.
 Blood Viscosity will be increase if blood flow r
ate decreases.
 Physics and Chemical
Properties of Blood
3. Specific Gravity （血液的比重）
Blood: 1.050-1.060
Plasma: 1.025-1.030
RBC: 1.090-1.092

RBC > Blood > Plasma

 Physics and Chemical
Properties of Blood
4. Plasma Osmotic Pressure

Osmotic pressure lies on the number of solu
te granule.
 Plasma osmotic pressure: 300 mmol/L

Plasma osmotic pressure = crystal osmotic pre
Osmosis
 Plasma Osmotic Pressure
Crystal osmotic Colloid osmotic
pressure pressure
Build up inorganic ions ( Na+ , protein
with Cl- , K+) & small orga
nic molecules
Role Balance water Balance water
between inside and between inside and
outside of the cell outside of the
vessel
Normal 285.5mOsm 1.5mOsm
value
 Plasma Osmotic Pressure

 Diffusion of water (solvent) across a

selectively permeable membrane, eg, ce
ll membrane.
 Important because large volume
changes caused by water movement
disrupt normal cell function.
Cell shrinkage or swelling ：
Isotonic: cell neither shrinks nor swells
Hypertonic: cell shrinks (crenation)
Hypotonic: cell swells (lysis)
 Iso-osmotic Solution & Isotonic Solution

Iso-osmotic Solution Isotonic Solution

definition have the same have the same
osmotic with osmotic with
plasma. plasma;
RBC can remain
normal shape in it.
Solute Can or can’t go Can go through cell
through cell membrane
membrane
example 0.9% NaCl, 0.9% NaCl
1.9% urea
 Section 2

Blood Cell
Red Blood Cell
Platelets
 Red Blood cell
(Erythrocyte)

 Function of RBC

 Properties of RBC

 Production of RBC and its regulation

 Function of RBC
 Transport
O2, CO2
 Buffering
– KHb/HHb
– KHbO2/HHbO2
– K2HO4/KH2PO4
– KHCO3/H2CO3
 Properties of RBC
1. Permeability
Permeate to O2, CO2, ure
a, Cl-, HCO3-

2. Reversible deformation
( 可塑性变形）
Volume (90 m3): Surface
area(140 m2)
Surface area in ball shape
d RBC is 100 m2
 Properties of RBC
3. Osmotic F
ragility
( 渗透脆性 ) ：
0.9% 0.8% 0.46% 0.34%
The resistance of R


BC to hypotonic
solution.
指红细胞在低渗盐溶液中发生膨胀、破裂和溶血的特性
，用于表示红细胞对低渗盐溶液的抵抗能力
 Properties of RBC
4. Suspension stability ( 红细胞的悬浮稳定性）
During a certain time RBC in blood can suspen
d relatively stably.
Erythrocyte sedimentation rate (ESR,
红细胞沉降率） : The rate at RBC settle o
ut of suspension in plasma is called ESR.
 Measurement of ESR
Put anti-coagulated blood
in vertical tube, then
RBC will sink slowly for
its larger density.

ESR is expressed by
RBC sinking distance
during the first hour.

 Normal value by Wei’s method:

male,0~15mm/h; female,0~20mm/h
 Factors that affect ESR
in plasma

 Rouleaux formation ( 叠连形成）

 RBC sticks to each other with concave.
 Rouleaux formation increase ESR.
 The reason lies in plasma change but not RBC
itself.
 Factors that affect ESR
in plasma
 Any plasma change that will affect RBC su
rface properties will make RBC tend to stick
together.
 Fibrinogen, some immunoglobulins increas
e ESR.
 Increased ESR is a measure of the acute ph
ase response to a challenge that may be infec
tive, ischemic, traumatic or malignant
 ESR increases with age
 Production of RBC
 Materials for production of RBC
 Iron( 铁 ):
• Old RBC release 95%; Absorption 5% .
• The amount of iron absorption is adjusted by
intestinal epithelium, depending on the state
of the body’s iron balance.
• The more iron in the body, the more in the i
ntestinal epithelium, and the less new iron wi
ll be absorbed.
Proteins( 蛋白质 ):intake with food

Vitamin B12 (cobalamin 钴胺素 ): The absorption

of vitamin B12 from gastrointestine requires a prot
ein, intrinsic factor ( 内因子 ) secreted by gastric pa
rietal cell.
Be essential for the formation of DAN.

Folic acid( 叶酸 ):It is essential for the formation

of DAN, and hence for normal cell division since f
olic acid is required for synthesis of the pyrimidine
thymine ( 胸腺嘧啶核苷酸）
 Production of RBC
The site of RBC production
 Bone marrow ( 骨髓 ), specifically the “re
d” one.
 In adulthood it is different from childhood.
child: liver, spleen
 The Factors That Affect the
Production of RBC
 Iron intake not enough or lose too much
 Vitamin B12 absorption not enough
Stop DNA
 Intrinsic factor deficiency synthesis
 Folic acid absorption not enough
 Regulation
of RBC Formation
 In adult, there are total 251012 of RBC. The
average life span of an erythrocyte is about 120
days.
 Almost 1 percent of the body’s erythrocytes are
destroyed and must be replaced every day (100
billion cells/day)
 A process of negative feedback controlled by
erythropoietin (EPO).
 Negative Feedback Cycle

Stem cellsBFU-ECFU-Eerythrocyte progenitor cells

The number of
EPO RBC

Cells that generate EPO

In kidney
 O2
High altitudes
Chronic lung disease
 O2 deliver to kidneys Blood loss (acute or
chronic) etc.
 Platelets
 Overview
 Platelet (or thrombocyte): mean diameter
2~4m, smallest formed elements in blood
 Formed in marrow: 2/3 traveled with
blood in vessel, 1/3 stored in liver
 Life time 7 to 14 days
The role of platelets

 Promote coagulation
 Involving hemostasis
 Be nutrient to endotheliu
m of capillaries
 Properties of platelets
 Platelet adhesion
 Platelet aggregation
 Platelet release
 Platelet contraction
(to compact clot mass)
 Platelet absorb
coagulation factor Platelet activation
 Platelet adhesion
 Platelet can’t adhere to the surface of normal en
dothelial cells.
 Platelet adhesion needs:
1. Glycoprotein (GP, especially GPIb) on platel
et membrane;
2. Subendothelium tissue (especially collagen fi
bers)
3. Von willebrand factor (vWF) in plasma.
 collagen fibers—vWF—GPIb
 Reagents That Induce
Platelets Aggregation
 ADP
 Thromboxane A2 (TXA2, ，血栓素 A2)
 Collagen ( 胶原 )
 Thrombin ( 凝血酶）
 Pathologic ： bacteria, virus, immune
complex, durg, etc.
Reagents That resist
Platelets Aggregation

In normal situation,
TXA2 and PGI2
keep a dynamic
balance.
 Platelet release
 Platelet can release 5HT, PG, histamin, ATP,
PF3, TXA2, etc.
 The reason that can induce platelet aggregati
on can almost also induce its release.
 Many of the released substances can promote
platelet activation, aggregation and accelerate
coagulation.
 Section 3

Hemostasis
Coagulation
Fibrinolysis
Hemostasis
 Hemostasis
 Definition: the process the body uses to
stop the flow of blood when the vascular
system is damaged.
 STEPS:
1. Vasoconstriction
2. Platelet plug
3. Blood Coagulation
 Hemostasis Is a Two
Stage Process
• The primary stage is characterized by
vascular contraction, platelet adhesion
and formation of a soft aggregate plug.
• The secondary stage is responsible for
stabilizing the soft clot and maintaining
vasoconstriction.
Hemostasis Process
 Endothelial cell
injury and
Subendothelium
uncover
 Platelet activated;
Vascular
constriction;
Coagulation system
activated
 Platelet plug and
fibrin formation
 Bleeding time
 Definition: Pinpoint pierce into the
earlobe or fingertip, then the bleeding
lasting time is Bleeding time (BT).
 It is a measure about the function of
platelet.
 Normal value of BT: 1 to 3 min
 Coagulation & Fibrinolysis

 Blood coagulation (clotting)

 Anticoagulant
 Fibrinolysis
 Blood Coagulation
 Definition
 a process in
which liquid blood
is changed into a
semisolid mass (a
blood clot)
 Coagulation Factors
Factor Name Plasma
half-life (h)

I Fibrinogen 纤维蛋白原 72 - 96

II Prothrombin 凝血酶原 60
III Tissue Factor or thrombopl -
astin 组织因子
IV Ca++ -
V Proaccelerin 前加速素 15
VII Proconvertin 前转变素 5
VIII Antihemophilic A factor 抗 10
血有病因子
Factor Name Plasma
half-life
(h)
IX Antihemophilic B factor or Christmas f 25
actor 血浆凝血激酶
X Stuart or Stuart-Prower factor 40
XI Plasma thomboplastin antecedent 血 45-65
浆凝血激酶前质子
XII Hageman factor, contact factor 接触 60
因子
XIII Fibrin stabilizing factor 纤维蛋白稳 150
定因子
Prekallikrein factor 前激肽释放原
High-molecular-weight kininogen 156
高分子激肽原
Three essential steps of Clotting
 Formation of prothrombinase complex
 Prothrombin (FII) activated
 Formation of fibrin (Ia)
Formation of prothrombinase complex
（ FXa-FVa-Ca2+- 磷脂复合物）

II IIa

I Ia
 Formation of prothrombinase
complex

 Intrinsic Pathway
 Extrinsic Pathway
 Intrinsic pathway interconnecte with ex
trinsic pathway
 Intrinsic Pathway
 Intrinsic Pathway is defined as a cascade t
hat utilizes only factors that are soluble in the
plasma.
 Surface activation ： a process from
FXII combining with alien substance
to FXIa formation.
 FXIa activate FIX in the present of Ca2+
, then FIXa combine with FVIIIa to form
tenase bomples, which can activate FX.
 Contact with the collagen und
er subendothelial cell
Surface activation
 Extrinsic Pathway

 Extrinsic Pathway triggered by tissue dam

age, which cause the release of factor III
（ Tissue Factor ）
 FIII combine with FVII to form FVIIa-FII
I complex, which activate FX.
 Factor III are distributed in various tissue,
especially lung, brain, placenta ( 胎盘 ).
 Contact with the collagen und
er subendothelial cell
Surface activation
 Intrinsic pathwa
y interconnect wi
th extrinsic path
way
TF-VIIa complex
activate FIX.
 Amplification of
coagulation
FIIa activate FXI.
Formation of fibrin
 Summary

 The classical model of blood

coagulation involves a series (or
"cascade") of zymogen activation
reactions.

 Coagulation can be initiated via the

“intrinsic” or “extrinsic” pathway.
 Summary
 Both the intrinsic and the extrinsic pathways
proceed through a common pathway by
forming activated factor X (factor Xa-Va-
Ca2+)

 Amplification of coagulation reactions.

 The intrinsic and the extrinsic pathways

are highly interconnected.
 Intrinsic and the extrinsic
pathways interconnect

Evidences:
1. The TF-VIIa complex activates no
t only factor X but also factor IX of t
he intrinsic pathway.
Factor VIIa activated by TF not only activates factor
X but also activates factor IX in the presence of TF,
providing a connection between “extrinsic” and “intr
insic” pathway.
Evidences:
2. Patient with severe factor VII deficiency
may bleed even though the intrinsic path
way is intact.

3. The severe bleeding associated with defici

encies of factor VIII or IX would not be e
xpected if the extrinsic pathway alone we
re sufficient to achieve normal hemostasis
.
 More recent evidence has shown
that the pathway are not, in fact,
redundant but are highly
interconnected.
Factors That Affect Coagulation

 Temperature: T  coagulation 
 Rough surface
 Chemical ： Ca2+, chelates ( EGTA
, EDTA, Sodium citrate, Potassium c
itrate ）
 Biochemical reagents, vitamin K
 Anticoagulative System
 The reasons that blood can flow through vessels
fluently are as follows:
– There is no injury in blood vessel.
– Most of clotting factors are present in blood
with inactive state.
– There are some anticoagulants (Compounds that
do not allow blood to clot) exist in blood (most
important).
 Anticoaglants
 Endogenous Inhibitors of Clotting
1. serine proteases inhibitor 丝氨酸蛋白酶抑
制物
 Antithrombin (major, 抗凝血酶 III): pr
imarily neutralizes factor Xa and thrombi
n, in addition to inhibiting most active of
the clotting system.
 2 -macroglobulin (2- 巨球蛋白）
 1 -proteinase inhibitor( 1- 蛋白酶抑
制物）
2. Protein C system
 Protein C is another plasma protein that
limits clotting by being activated by throm
bin to proteolytically inactivate proacceleri
n (V) and antihemophilic factor (VIII).

 Thrombomodulin ( 凝血酶调制素 ) speci

fically binds thrombin (II) so as to convert
it to a form with decreased ability to cataly
ze clot formation.
 Anticoaglants
 Exogenous Inhibitors of Clotting
1. Heparins (肝素）

2. Coumarins( 香豆素） competitive inhi

bitors of vitami
n K in the biosy
3. 1,3-indanedions   nthesis of prothr
ombin
1. Heparin forms a high-
affinity complex with
antithrombin (AT-H).

2. The formation of AT-

H complex greatly inc
reases the rate of inhib
ition of two principle
procoagulant protease
s, factor Xa and thro
mbin (II).
4. Tissue factor pathway inhibitor (TFPI, 组织
因子途径抑制物 , or lipoprotein-associated c
oagulation inhibitor ）
 TFPI in vivo is thought to be synthesized m
ainly by endothelial cell.
 TFPI inhibits the activation of the extrinsic
coagulation cascade through its ability to in
hibit factor Xa and combine with FVIIa-TF
complex.
 VII Ca2+
TF—VIIa—TFPI—Xa
TF Complex of inhibitor
Ca2+
(2)
Ca2+
TF—VIIa
TF —Xa
IX IXa (1)

X Xa TFPI

II IIa
Catalysis Convert
 Fibrinolysis

Breakdown of Blood Clots

 Fibrinolysis
 Fibrinolysis
– Fibrinolysis leads to the breakdown of fibri
n clots (blood clots) and is caused by the act
ion of several enzymes

– Whenever fibrinolysin is formed in a blood

clot, it can cause lysis of clot and also destr
uction of many clotting factors, thereby cau
sing hypocoagulability of the blood.
 Kallikrein ( 激肽释放酶 )
Endothelium Cl inhibitor
( 内皮细胞）
uPA Urokinase
( 尿激酶原 )
tPA
PAI-1
( 内皮细胞）
Plasminogen Plasmin ( 纤溶酶 )
( 纤溶酶原） 2-antiplasmin
2-macroglobulin
Fibrin
Fibrin (or
degradation
fibrinogen
products
)
Catalysis; Convert; Inhibit. PAI-1 纤溶酶原
激活物抑制剂
 Section 4

ABO Blood Types

 The most well known and
medically important blood
types are in the ABO group. 
 Discoverer
• Karl Landsteiner d
iscovered ABO bl
ood type in 1900 a
nd 1901.
• In 1930, he receive
d the Nobel Prize f
or this discovery.

Karl Landsteiner
(1868-1943)
 ABO blood type

 There are four types: A, B, AB,

and O decided by the antigens
(Antigen A/B) on RBC
membrane. 

 There are two antigens and two

antibodies that are mostly
responsible for the ABO types. 
Antigen and Antibody in ABO Blood Type

ABO Antigen  Antigen  Antibody Antibody

Blood A 
Type B  anti-A anti-B
A yes no no yes

B no yes yes no

O no no yes yes

AB yes yes no no  

 Genetic Inheritance Patterns
 ABO blood types are inherited through
genes on chromosome 9
 Do not change as a result of environmental
influences during life.
 An individual's ABO type is determined by
the inheritance of 1 of 3 alleles (A, B, or O)
from each parent.
Genotype AA AO BB BO AB OO
Phenotype A A B B AB O

 The alleles must be two of A, B and O to

form a person’s Genotype.

 Both A and B alleles are dominant over O

 Parent Alleles
A B O
                 

AA AB AO
A
(A) (AB) (A)
AB BB BO
B
(AB) (B) (B)
AO BO OO
O
(A) (B) (O)

The possible ABO alleles for one parent are in the top
row and the alleles of the other are in the left
column. Offspring genotypes are shown in white.
Phenotypes are red.
Unagglutinated blood smear Agglutinated blood

Sometimes when the blood of two people

is mixed together, it clumps or forms
visible islands in the liquid plasma--the red
cells become attached to one another.  
This is agglutination.
Antibodies seeking Antibodies agglutinating
specific antigens red cells
Anti B Anti A Anti-A-B (Serum)
ABO Blood Type
Diagnosis
B

AB

O
 Blood Transfusion

 Advances: transfusion of blood

components

 Cross-match test
1. Main test: Donor’s RBC + Recipient’s serum
2. Subordination test: Donor’s serum +
Recipient’s RBC