Chapter 3 BLOOD PHYSIOLOGY

Wang Guoqing
Department of Physiology, Medical School, Soochow University, Suzhou 215123, China

E-mail:wangguoqing@suda.edu.cn Tel:0512-62096158; 13506212030

I. II. Physical and chemical characteristics of blood III. Blood Cells 1. Hemopoietic process and hemopoietic stem cells 2. Hemopoietic microenvironment 3. Erythrocyte Physiology 4. Leukocyte Physiology 5. Platelet or Thrombocyte Physiology IV. Physiological Hemostasis 1. Endocrine functions of vessel endothelial cells 2. Physiological Characteristics of Platelet 3. Blood Coagulation 4. Fibrinolysis V. Blood Group 1. RBC Agglutination 2. ABO blood group system 3. Rh blood group system 4. Relation between blood volume and clinic 5. Principle of Transfusion and Cross-match test

What will we discuss in this chapter? (Outline) Blood composing

Outline

Blood and Internal Environmental Homeostasis

Blood is that part of extracellular fluid within the cardiovascular system Blood forming During animals evolution, extracellular fluid was gradually shaped from the age-old time with ocean which was mainly salty solution. At last, extracellular fluid was differentiated into plasma and interstitial fluid and blood came from plasma and cells. The role of blood in internal environmental homeostasis Blood, the most active component in extracellular fluid, display functions as follows:

(1) (2) (3) (4)

transportation; pH value buffer; temperature or thermal maintenance; immunity and defence

I. Blood composing

Blood composing: plasma + blood cells Hematocrit: blood cells occupies the percentage of total blood volume. normal value male: 40-50% female: 37-48% newborn: 55%

Blood component (summing-up)

Terminology and normal value

Chemical component of plasma

Water: > 90% Small molecule: 2%, it is electrolytes, nutriment, metabolic products, hormone, enzyme,etc. Protein: 60-80 g/L, plasma protein include albumin (40-50 g/L), globulin (20-30 g/L,1-, 2, -, - ) and fibrinogen. Most of albumin and globulin made from liver. A/G and clinic. Function of plasma protein: (1) transportation, (2) nutrition, (3) forming colloid osmotic pressure, (4) coagulation and anticoagulation, (5) pH value buffer, (6) immunity (globulin)

Chemical component of plasma

H 2O 90 - 91%
Plasma
Interstitial fluid Intracellular fluid

Na+ ClCa++ K+ PO4Protein

142 104 2.5 4.3 2 14

145 117 2.4 4.4 2.3 0.4

12 4 <0.001 139 29 54
(Unitmmol/L)

II. Physical and chemical characteristics of blood

Specific gravity: total blood (1.050-1.060) more influenced by red blood cells; plasma (1.025-1.030) more influenced by plasma protein; RBC (1.090-1.092) more influenced by Hb. Viscosity: Blood relative viscosity (4~5) mainly depends on the numbers of red blood cells. Plasma relative viscosity (1.6~2.4) is mainly involved in plasma protein Plasma osmotic pressure is 300 mmol/L or 770kPa (1) Crystal osmotic pressure results from NaCl and modulates water distribution between inside and outside of cells. (2) Colloid osmotic pressure results from albumin and regulates water distribution between inside and outside of capillary. Plasma pH value is about 7.35~7.45, and usually buffer systems are NaHCO3/H2CO3 (20:1), protein salt/protein, Na2HPO4/ NaH2PO4, Hb salt/Hb, HbO salt/ HbO2, K2HPO4/ KH2PO4, KHCO3/H2CO3, etc [lungs and kidney mainly regulate Plasma pH value ].

Osmosis and Osmotic Pressure

Osmosis is the movement of water down its concentration gradient. Osmosis is determined by the number of impermeable molecules. Osmotic pressure is the force drawing water down its concentration gradient.

Osmosis and Osmotic Pressure

A B

Water

[Water] > [Water] [Salt] < [Salt] Osmotic Pressure < Osmotic Pressure Osmosis is the movement of water from a high concentration to a low concentration. In this illustration, two compartments (A and B) are separated by a semipermeable membrane (broken vertical line). The water concentration in compartment A is greater than the concentration in compartment B because of the presence of salt (X) in B. Therefore, water will move down its concentration gradient from A to B. The force needed to prevent this water movement is called osmotic pressure.

Tonicity

The tonicity of a solution refers to the effect of the solution on cell volume. A hypertonic extracellular solution is one in which the water concentration is less outside the cell than inside; water leaves the cell; cell volume decreases. An isotonic extracellular solution is one in which the water concentration is the same inside and outside the cell; no water movement; cell volume does not change. A hypotonic solution is one in which the water concentration is greater outside than inside the cell; water enters the cell; cell volume increases. An isosmotic solution may not be an isotonic solution if the particles are permeable to the cell membrane.

III.Blood Cells
Blood cells are erythrocyte (red blood cell, RBC), leukocyte (white blood cell, WBC) and thrombocyte (platelet, P).

Blood Cells

The forming processes of erythrocyte (red blood cell, RBC), leukocyte (white blood cell, WBC) and thrombocyte (platelet, P) originating from hematopoietic stem cells are hemopoiesis. Transfer of blood cells forming place: yolk sac hemopoiesis (early embryo period) liver and spleen (second embryo month) marrowand liver, spleen (after fourth embryo month) marrow (fetus birth time) and liver, spleen as complementary role. During adulthood (after 18), red marrow (flat bones, e.g. vertebra,ilium, sternum, rib, skull and long bone ending) rather than yellow marrow has hematopoietic functions.

1. Hemopoietic process and hemopoietic stem cells

Hemopoietic process
Stage one: Hemopoietic stem cells

self renewal, steady numbers, active differentiation.

Stage two: committed progenitors directional differentiation (CFU-GEMM, CFU-E, CFU-

GM, CFU-MK, CFU-TB). [CFU: colony- forming unit

Stage three: precursors morphologic occurrence of various original blood cells.

Hemopoietic stem cells

Basic characteristics Self renewal in high degree, constant from young to old age. Multi- directional differentiation Large potential proliferation, Hemopoietic stem cells produce about 11011 blood cells releasing to blood for use. Surface sign According to CFU (colony forming unit), using fluorescence-activated cell sorting (FACS), its main surface sign is CD34+CD38-Lin-and CD34-CD38-Lin-. Note CD: cluster of differentiation of antigen on the white blood cells; Lin: systemic specific antigen on the hemopoietic cells.

2.Hemopoietic microenvironment

Hemopoietic microenvironment: It includes stromal cell secreting extracellular matrix (ECM), multihemopoietic regulating factor, hemopoietic nerves and blood vessels. Stromal cells in the marrow come from fibrocyte, reticulocyte, endothelial cell, ectoblast cell, monocyte, engulfing cell, osteoblast and osteoclast. Stromal cells supply two material: one is soluble hemopoietic growth factor, another is membrane-combined adhesive molecule. Extracellular stroma synthesized and secreted by marrow stromal cell filling cellular interstice contains big molecules, such as collagen (typeI, II, III, IV), glycoprotein (fibronectin, laminin, hemopoieticnectin ) and protein amylose (sulfate cartilagetin, sulfate heparin, hyaluronic acid and sulfate dermatin, etc). Hemopoietic cells must adhere to stromal cell and is in the hemopoietic microenvironment for survival.

Hemopoietic process

Hemopoietic process

Hemopoietic process

3.Erythrocyte Physiology

Shape and number of red blood cells (RBC) Shape of RBC: like biconcave disc

Its diameter is about 7~8 m, peripheral thickness about 2.5 m, central thickness about 1 m and cubage about 90 m3.

Reason for shape of RBC

biconcave disc like

Erythrocyte Physiology
Number of RBC: It is most numbers in the blood. Normal value about RBC Male adult, 4.5~5.51012/L; average, 5.01012/L Female adult, 3.8~4.6 1012/L; average, 4.21012/L Newborn, 6.01012/L Protein within RBC is hemoglobin (Hb). Hb in male adult, 120~160 g/L; Hb in female adult, 110~150 g/L; Hb in newborn (within 5 days), 200 g/L Pregnant female, numbers of RBC and Hb are relatively less (because of more plasma). Dweller lived in plateau, numbers of RBC and Hb are relatively more (because of compensation for anoxia).

Physiological Characteristics and Functions of RBC

Characteristics of RBC Permeability: semipermeable membrane, gas and urea freely passing through, negative ions easily in or out of RBC, and positive ions not. There are NaK ATPase as pump on the membrane of RBC and low-temperature-stored plasma easily has high kalium. Why? Plasticity and metamorphose:

Plasticity and metamorphose depend on: 1) surface area-cubage ratio, 2) viscosity of Hb, 3) membrane elasticity and viscosity.

Physiological Characteristics and Functions of RBC

Characteristics of RBC Suspension stability: it cab be described by erythrocyte sedimentation rate (ESR) which is RBC descending distance per hour and suspension stability is inverse proportion to ESR. Normal value of ESR: male, 0~15 mm/h; female, 0~20 mm/h. ESR and clinic: some diseases bring about rouleaux formation (mainly involved in plasma component, e.g. globulin, fibrinogen, cholesterol) and speed up ESR.

Physiological Characteristics and Functions of RBC

Characteristics of RBC Osmotic fragility: Changes in RBC put into lower osmotic salty solution. Osmotic fragility of aged RBC is large and easily results in rupture (hemolysis and ghost cell). Isosmotic solution, e.g. 0.85% NaCl, 1.4%NaHCO3, 5% glucose, etc. Isotonic solution, e.g. 0.85% NaCl Isosmotic solution does not equal to isotonic solution. Isosmotic solution, isotonic solution and clinic

Physiological Characteristics and Functions of RBC

Functions of RBC

RBC can be used for transportation of O2 and CO2 in the blood. RBC can be served as pH buffer.

Erythropoiesis

Hemopoietic material for erythropoiesis: iron (Fe++) and protein, [reason for anemia] Influencing factors of RBC maturity: Vitamin B12 and folic acid (DNA metabolism), [clinic relation] Process of erythropoiesis:
Hemopoietic stem cellsmulti systemic hemopoietic progenitor cellsRBC-committed progenitor cells (BFU-ECFUE)original RBC earlier infantile RBCmedium-term infantile RBCterminal infantile RBCreticular RBCmature RBCblood for circulation.

This process requires 6~7 days. [mitosis several times] [apoptosis]

Place for Erythropoiesis

Main place for Erythropoiesis is bone marrow. Aother place is liver.

Regulation of Erythropoiesis

0.8% of total RBCs has self renewal, that is to say, 160106 RBC production every minute. Burst forming unit-erythroid, BUF-E, important to earlier erythropoiesis, depends on stimulation of burst promoting activity, BPA outside body. BPA made by leucocyte is a glycoprotein whose molecular weight is about 25000~40000 Colony forming unit-erythroid, CFU-E, important to terminal erythropoiesis, depends on erythropoietin, EPO which is also a glycoprotein, molecular weight, 34000, plasma concentration 10 pmol/L, half life 5 hours, increasing release when anoxia.

Regulation of Erythropoiesis

Life and breakage of RBC

Life-span: 120 days, about 4 months, each RBC circulates 27 km averagely in vessels, short life-span for aged RBC Breakage: places are liver, spleen and lymphatic node, and after breakage, Hb released from RBC immediately combine with plasma 2-globulin (Hb touched protein) which is taken in by liver for iron reuse.

Hb, very toxic if it get into blood, normally, it can be metabolized into bile pigment in liver.
Clinic relation.

4.Leukocyte Physiology Classification and numbers of Leukocyte

Number of Leukocyte (white blood cells, WBC): (4.0~10)109/L

Classification: It is granulocyte (neutrophil,

eosinophil, basophil), monocyte and lymphocyte.

Classification and numbers of Leukocyte

TABLE. Classification and normal value of Leukocyte
Absolute Value (109/L)
Total numbers of leukocytes 4.0~10.0
1~5 50~70 0.5~5 0~1 3~8 20~40
For Clinic Use

Percentage (%)

Neutrophil (bacilliform nucleus) 0.04~0.5 Neutrophil (foliiform nucleus) Eosinophil Basophil Monocyte Lymphocyte 2.0~7.0 0.02~0.5 0.0~0.1 0.12~0.8 0.8~4.0

Physiological Changes in Numbers of Leukocyte

Newborn: Number is higher, 15109/L, after birth 3 or 4 days to 3 months, being about 10109/L, mainly, neutrophil, 70%; secondarily, lymphocyte. Circadian changes: Number of WBC is more in the afternoon than in the morning. Food taking, ache and mood excitation: Number of WBC is remarkably higher. Heavy exercise and laboring: Increasing numbers, about 35109/L, return to original level after action stop. Terminal pregnancy of female: Numbers changes in 12~17109/L, and during parturition, 34109/L, and after parturition 2~5 days, number return to original level.

Physiological Characteristics and Functions of WBC

Terminology Diapedisis: Metamorphosed WBCs pass through vessel wall getting into interstitial fluid. Chemotaxis: It is a process that WBCs shift to some chemical material (metabolic production, antigen-antibody complex, bacteria, toxin, etc). Phagocytosis: It is a process that WBCs enclose and engulf exotic or extraneous material, and use intracellular enzyme digesting them.

WBC Diapedisis

Blood Vessel

Metamorphose

Physiological Characteristics and Functions of WBC

Neutrophil

Another name, polymorphonuclear, PMN, 6~8 h in the vessels, diapedisis, chemotaxis and phagocytosis (using its hydrolyzed enzyme) Function: It plays a very important role in nonspecific cellular immunity system which is against pathogenic microorganism, such as bacteria, virus, parasite, etc. Clinic relation: Number of neutrophil greatly increase occurring in acute inflammation and earlier time of chronic inflammation. number decrease of neutrophil will result in poor resistibility and easily suffering from infection.

Physiological Characteristics and Functions of WBC

Eosinophil
Circadian changes: Its number is lower in the morning
and higher at night. Function:

1. It limits and modulates the effects of basophil on fast

allergic reaction. 2. It is involved in immune reaction against worm with

opsonization.
Clinic relation: Its number increase when person suffers from parasite infection or allergic reaction.

Physiological Characteristics and Functions of WBC

Basophil
Circulatory time: 12 hours Basogranules contain heparin, histamine, chemotactic factors and chronic reactive material for allergic reaction. Function: It is also involved in allergic reaction. 1. Heparin serves as lipase cobase and speeds up fatty decomposition. 2. Histamine and chronic reactive material increase permeability of capillary and contract bronchia smooth muscle, and result in allergic reaction such as measles, asthma. 3. Eosinophil chemotactic factor A released by basophil can attract eosinophil collection and modify eosinophil function.

Physiological Characteristics and Functions of WBC

Monocyte
Its body is large, diameter about 15~30 m without granule Function: 1. It contains many nonspecific lipase and displays the powerful phagocytosis. 2. As soon as monocytes get into tissue from blood , it change name called macrophage activating monocyte- macrophage

system to release many cytokins, such as colony stimulating

factor (CSF), IL-1, IL-3, IL-6, TNF, INF-, ,etc. 3. Cytokins induced by monocyte may modulate other cells growth. 4. Monocyte- macrophage system plays a very important role in specific immune responsive induction and regulation.

Physiological Characteristics and Functions of WBC

Lymphocyte
Classification: It can be separated into T- Lymphocyte and B- Lymphocyte. Function: 1. Lymphocytes serve as a nuclear role in immune responsive reaction. 2. T- Lymphocytes involved in cellular immunity. 3. B- Lymphocytes involved in humoral immunity. Clinic relation: Numbers increase of lymphocytes occur in

Leukopoiesis, Regulation and Breakage

Birth place: bone marrow, originating from hemopoietic stem cells, and leukopoiesis process is similar to RBC. Leukopoiesis, differentiation and growth are influenced by hemopoietic growth factor, HGF which are glycoprotein secreted by lymphocyte, monocytemacrophage, fibrous cell and endothelial cell. Colony stimulating factor, CSF, such as GM-CSF, G-CSF, M-CSF, Multi-CSF (IL-3) also influence Leukopoiesis. Life span: several hours to 3 or 4 days. Leukocyte breakage: site are liver, spleen and lymphatic node. Pus or purulence forming

5.Platelet or Thrombocyte Physiology

Shape: Biconvex disk like, diameter about 2~4 m, average cubage 8 m3. Complicated structure: under the electronic microscope, there are -granule, dense body, lysin peroxide enzyme, opening tubular system, dense tubular system, canaliculus,etc. Dense body: It contains ADP, ATP, 5-HT, Ca2+, epinephrine,etc. Source: Platelet comes from megakaryocyte fractionlet release in the marrow.

Normal Value and Function of Platelet

Normal value: 100109 ~ 300109, range from 6%~10% Normal changes: more number in the afternoon than in the morning, more in winter than in spring, more in the venous blood than capillary, after sport, pregnacy. *Functions: 1. It maintains capillary endothelial cells smooth and integrated (repairing endothelium and providing nutrition). 2. It is involved in physiological hemostasis. Platelet and clinic relation: decrease of platelet, abnormal immune reaction, will results in hemorrhage or bleeding, purpuric symptom.

Platelet Forming and Regulation

Platelet forming: Birth place is bone marrow, originating from hemopoietic stem cells, and differentiating into burst forming unitmegakaryocyte, BFU-MK, then continuously into CFU-MK, and into megakaryocyte, demarcation membrane system, DMS, into fractionlet release to the blood requiring 8~10 days. (one megakaryocyte can produce 200~7700 platelet). Regulation: Protein, Mpl, expressed by c-mpl (oncogene) exists in CD34+ located at hemopoietic stem cells/ committed progenitors, megakaryocyte and platelet, found by Methin in 1993, and its ligand named thrombopoietin, TPO was discovered in 1994 which promoted hemopoietic stem cells differentiating into megakaryocyte as hemopoietic stem cells positive regulating factor.

Life- Span and Breakage of Platelet

Life-span: Averagely, 7~14 days in the blood. It can be consumed when it displays physiological functions. Breakage: Aged platelet can be processed by phagocytosis in liver, spleen and lymphatic node.

IV. Physiological Hemostasis

*Definition: The process from vessel bleeding to automatic hemostasia.

*Bleeding time: The time from vessel bleeding to automatic hemostasia. Normal time is 1~3 min and it is longer when platelet decrease. Process of hemostasis: 1. Blood vessel contraction or convulsion (induced by neuroreflex; 5-hydroxytryptamine,5-HT; thromboxane A2, TXA2; endothelin, ET ) 2. Platelet thrombosis forming (made by platelet adhesion, aggregation, release and contraction)

3. fibrin, clot forming and maintenance (made by blood coagulation activation)

Physiological Hemostasis

1.Endocrine functions of vessel endothelial cells

Material related to hemostasis are basal membrane, collagen (III, IV), microfibril, elastin, laminin, ectonectin, fibronectin, von Willebrand factor (vWF), protein enzyme, protein enzyme inhibitor, adhesive amylose, etc. Anticoagulative material: They are prostacyclin (PGI2), endothelium-derived relaxing factor (EDRF or nitric oxide, NO), tissue-type plasminogen activator (tPA), uPA, ADPase, ATIII, heparin sulfate, protein C, thrombomomodulin (TM), plasminogen activator (PA). Promoting coagulative material: Tissue factor, vWF, blood clotting factor V, plasminogen activator inhibitor (PAI-1, PAI-2, ATIII), TNF, interleukin-1 (IL-1). Vessel constricting and relaxing modulators: endothelin1 (ET-1), EDRF (NO), PGI2, etc.

Roles of Vessel Endothelial Cells in Physiological Hemostasis

Roles are close related to its endocrine functions
Vessel endothelium serves as barrier between underendothelial structure (namely, collagen) and blood. As soon as collagen expose to blood, hemostasis of platelet is immediately activated to form thrombus blocking wounded vessels. Platelet activation can releases constrictive factors (TXA2, ET-1, 5-HT, etc) making vessel convulsion, lasting about 60 sec. Stimulated vessel endothelial cells release coagulative factors and Promoting coagulative material to realize, speed up blood coagulation. At the same time, cells also release anticoagulative factors and fibrinolysis material to modify blood coagulation.

Inactive Platelet
Under the electronic microscope

Activated Platelet for Hemostasis

Under the electronic microscope

2.Physiological Characteristics of Platelet

Thrombocyte adhesion: its membrane glycoprotein (GP, GPIb/IX and GPIIa/IIIb), collagen (underendothelial structure), vWF (plasma component), fibrinogen are involved in adhesion. Mechanism: Exposed collagen+vWF vWF changes platelet membrane glycoprotein+changed vWF Thrombocyte adhesion. Thrombocyte aggregation: induced by physiological factors such as ADP, thromboxane A2 (TXA2), epinephrine, 5-HT, histamine, collagen, thrombin, prostacyclin,etc and by pathological factors like bacteria, virus, immune complex, drugs, etc. The process can be separated into two phases: phase one is reversible aggregation and phase two irreversible aggregation. Two phases require Ca2+, fibrinogen and energy consumption. Mechanism : Various factors+corresponding receptors on the platelet changes in the second messenger within platelet cAMP, Ip3, Ca2+, cGMP platelet aggregation. Thrombocyte release: ADP, ATP, 5-HT, Ca2+ released from dense body, and -platelet globin, PF4, vWF, fibrinogen, PFV, PDGF, thrombin sensitive protein from -granule, and acid protein hydrolyzed enzyme, tissue hydrolyzed enzyme from lysosome. Thrombocyte contraction: Loose platelet thrombus could turn into compact platelet thrombus by Ca2+ release and cytoskeleton movement (filament/canaliculus) within platelet.

Roles of Platelet in Hemostasis

Activation of platelet: Stimulus brings about thrombocyte adhesion, aggregation, release and contraction. Loose platelet thrombus forming: First phase of hemostasis. Blood coagulation activation by platelet: Fibrin net forming, second phase of hemostasis. *Roles of platelet in hemostasis: 1. Activated platelets supply lecithoid (phospholipid) surface for blood clotting factor and involve in activating factor X and prothrombin. 2. Surface of platelet membrane combine with many blood clotting factor, such as fibrinogen, FV, FXI, FXIII to speed up coagulation. 3. Activated platelets release -granule which contains fibrinogen to intensify fibrin forming and blood coagulation. 4. Activated platelets contract clot with its contractive protein to solidify blood coagulation.

Two Phases of Physiological Hemostasis

First Phase

Second Phase

Mechanism1 of Platelet in Hemostasis

Mechanism2 of Platelet in Hemostasis

3.Blood Coagulation Blood Clotting Factor

Definition: The process of blood flow from flowing liquid to gel or gelatin. Serum: Light yellow fluid after blood coagulation. Difference between serum and plasma mainly consists in no fibrinogen in serum. Blood coagulation is a series of complicated biochemical reactions with various enzymes. Blood clotting factor: Material which are directly involved in blood coagulation. There are 12 factors named Roman numerals, except Ca2+, phospholipidother factors being protein, and except FIII (TF), others are in fresh plasma synthesized by liver with VitK . Blood clotting enzymes have two type: inactive and activated type [FII, FVII, FIX, Fx, FXI, FXII, FXIII].

Blood Clotting Factor

Factor Name Plasma Concentration I Fibrinogen 3000 II Prothrombin 100 III Tissue factor IV Ca2+ 100 V Proaccelerin 10 Proconvertin 0.5 Antihemophilic factor,AHF 0.1 Plasma thromboplastic 5 component,PTC(Christmas factor) Stuart-Prower Factor 10 Plasma thromoboplastin 5 antecedent,PTA Contact factor or Hageman factor 40 XIII Fibrin-stabilizing factor 10 - High-molecular weight 80 kininogen,HMW-K - Prekallikrein,Pre-K or Fletcher factor 35 Synthesizing site Liver Liver (with Vit K) Endothelial cell Endothelial cell, platelet Liver (with Vit K) Liver Liver (with Vit K) 4~5 d 3d 12~15 h 4~7 h 8~10 h 24 h Half life Chromsome site 4 11 1 13

Liver (with Vit K) Liver

Liver Liver, platelet Liver Liver

2d 2~3 d
24 h 8d -

13 4
5 6,1 3 4

Blood Coagulation

Intrinsic pathway of blood coagulation: All blood clotting factors involved in blood coagulation come from blood. Eyewinker surface with negative charges (collagenin) on the endothelium of blood vessel activates blood FXII as beginning of coagulation named surface activation. Extrinsic pathway of blood coagulation: Stimulus activates tissue factor (FIII) as beginning of coagulation. Extrinsic pathway of blood coagulation is faster than intrinsic pathway of blood coagulation because its steps are more simple. *Basic steps of blood coagulation [typical positive feedback]: Prothrombin activator forming [FXa-Va-Ca2+-phospholipid] Step 1

Prothrombin

thrombin

Step 2

Fibrinogen fibrin (clot) Step 3 Hemophilia A, B, C in the clinic results from deficiency of FVIII, FIX, FXI in the blood, respectively.

Process of Blood Coagulation

Extrinsic pathway

Tissue FactorTF
TF+ Ca2+ -TF a a-TF Ca2+ PL
PL: phospholipid

Intrinsic pathway

Eyewinker surface
Ca2+ HK a a S K PK

Ca2+ PL

a Ca2+ a PL a Ca2+ a PL

CL: cross linking fibrin

HK: high molecular weight kininogen S: Subendothelium PK: prekallikrein K: kallikrein

a
a Ca2+

CLa

Mechanism of Blood Coagulation

Cellular anticoagulative system: Liver cell and reticular endothelial cell could engulf blood clotting factor, tissue factor, prothrombin complex and soluble fibrin monomer. Humoral anticoagulative system: 1. Amino acid protease inhibitors in blood include antithrombin III, Clinhibitor, 1 antitrypsin, 2 antiplasmin, 2 huge globin, heparin coenzyme II, protease nexin-1 (PN-1) to combine with FIXa, FXa, FXIa, FXIIa and thrombin and then inactivate them for anticoagulation. Heparin can intensify functions of antithrombin III. 2. Protein C system are protein C (PC), thrombomodulin (TM), protein S and Protein C inhibitors. Main functions of PC consist in It inactivates FVa, FVIIIa with phospholipid and Ca2+; It blocks FXa combining with platelet phospholipid membrane to reduce prothrombin activation; It stimulates plasminogen activators release to trigger fibrinolysis; Protein S is a coenzyme of PC and greatly intensify functions of PC. 3. Tissue factor pathway inhibitor (TFPI) mainly coming from vessel endothelial cells inhibits FXa and inactivates FVIIa-TF complex to block extrinsic pathway of coagulation with negative feed back. 4. Heparin used in the clinic widely is due to It combines with antithrombin III to increase functions of antithrombin III; It stimulates vessel endothelial cell greatlu releasing TFPI and other anticoagulative material; It intensifies PC activation and stimulates vessel endothelial cell releasing plasminogen activators to increase fibrinolysis. [lower molecular weight heparin is less hemorrhage]

Anticoagulative system in blood

4.Fibrinolysis

Fibrinolytic system is involved in fibrinolysis, tissue repair and vessel rebirth. Two fibrinolytic systems: cellular one and plasma one. The former is leucocyte, macrophage, endothelial cell, mesothelial cell and platelet to engulf and digest fibrin. The latter is plasminogen activators (PA) and its inhibitors (PAI), plasminogen, plasmin. Basic steps:
Endothelial cells (Extrinsic
pathway ) (Urokinase, uPA)

Kallikrein (Intrinsic pathway) Cl-inhibitors uPAG

tPA Plasminogen

uPA

PAI-1

2-antiplasmin 2-huge globin

Plasmin
Fibrin dissolution

Fibrin or fibrinogen

Blood Coagulation and Fibrinolysis

Antifibrinolysis: Fibrinolytic Inhibitors and Its Functions

Main fibrinolytic inhibitors: They are plasminogen activator inhibitor type-1 (PAI-1, in platelet), 2antiplasmin (in liver), 2-huge globin, 1-antitrypsin, antithrombin III, alexin C1 inhibitor. PAI-1 synthesis and release: PAI-1 made by endothelial cell, smooth muscular cell, mesothelial cell, megakaryocyte is stored in platelet with inactive form. Some factors such as thrombin, IL-1, TNF, etc stimulate its release from platelet. PAI-1 function: It inhibits tPA (tissue-type plasminogen activator) limiting local fibrinolysis of thrombus. 2-antiplasmin characteristics: (1) Quick effect, (2) Inhibit plasminogen adhering to fibrin; (3) Combine with fibrin chain and block fibrinolysis Clinic relation: Innate deficiency of 2-antiplasmin often brings about serious hemorrhage.

V. Blood Group

History: ABO blood group system was firstly found by Landsteiner in 1901.
Definition for blood group*: Types of specific antigens on the blood cell.

Agglutination: Combination of the same antigen (or named agglutinogen, glycoprotein/glycolipid on the membrane of blood cell) and antibody (or named agglutinin, r-globin in serum) results in harmful immune reactions showing hemolysis.
Human leukocyte antigen, HLA have widespread distribution in the body and involves in immune repulsive reaction of organ transplant. Platelet antigens such as PI, Zw, Ko, etc may bring about fever heat when transfusion occur.

1. RBC Agglutination
Antigen-Antibody Harmful immune Reaction

Blood Coagulation

RBC Agglutination

Antigen of Blood Group

Antigen: Its genes are located at allele on euchromosome, namely, expressed gene.

Genotpye is genetic gene in blood group system and phenotype is antigen produced by corresponding genetic gene and amorph is noneffective allele. Genes in the blood system decide differential specific antigen on the membrane with control of enzymatic activity.

Antibody of Blood Group

Crude antibody: It is the unexposed antibody to correlative RBC, e.g., IgM in ABO blood group system which can not pass through placenta for the sake of big molecule.

Immune antibody: Various extraordinary RBC antigens (transfusion or parturition) sensitize lymphatic cells producing antibody such as Rh, Kell, Duffy, kidd, which belong to IgG (small molecule) and IgM (big molecule).

Blood Group of RBC

Number: 23 types, 193 antigens, more important blood groups are ABO, Rh, MNSs, Lutheran, kell, Lewis, duff, kidd, etc and all of them could result in hemolysis during transfusion. ABO blood group system:
Antigen on the RBC A B Antibody in the serum Anti-B Anti-A Anti-A+Anti-B

Blood group A B

AB
O

A+B

2. ABO blood group system

Antigen (agglutinogen) and antibody (agglutinin) in ABO blood subgroup system
Blood group
A B AB A1B A2B O A1 A2

Antigen on the RBC

A+ A1 A B A+ A1 +B A+B

Antibody in the serum

Anti-B Anti-B+ Anti-A1 Anti-A Anti-A1 Anti-A+Anti-B

Antigen of blood group Ushering material

ABH Antigen chemical structure in ABO blood group system

O(H)-antigen

A-antigen

B-Antigen

Galactose
Sugar

N-acetamide Glucose Glucose

N-acetamide galactose

Inheritance of ABO blood group

Inheritance: The A, B, H agglutinogen in ABO blood group system controlled by gene which is located at allele on No.9 chromosome (9q34.1-q34.2). Genotype and Phenotype:

Genotype and Phenotype in ABO blood group system

Genotype OO AA, AO BB, BO AB phenotype O A B AB

Inheritance of ABO blood group

Genetic relationship of ABO blood group
Parents blood group Offspring possible blood group Offspring impossible blood group

O O AA

O O, A

A, B, AB B, AB

AO
BB BO BA ABO ABA ABB

O, A
O, B O, B O, A, B, AB A,B A , B, AB A , B, AB

B, AB
A, AB A, AB ____ O, AB O O

ABAB

A , B, AB

Distribution of ABO blood group

Mid Europe: Type A 40%, Type O 40%, Type B 10%, Type AB 6%. America aborigines: Type O 90%. China Han nationality: Type A 31.31%, Type B 28.06%, Type AB 9.77%, Type O 30.86%. Other chinese minority is different. Bloog group can be used in research on anthropology

Mensuration of ABO blood group

Anti-B Serum Anti-A Serum Anti-A, B Serum

3. Rh blood group system

Rh antigen (Rh factor) is about 40 kinds and Rh factors related to clinic are D, E, C, c, e and most important is D antigen. Membrane of RBC has D antigen meaning Rh Positive, otherwise, Rh negative. Most of people (99) are Rh Positive and less than 1% persons are Rh negative. Rh blood group characteristics: Immune antobody and incomplete antibody, IgG; while ABO blood group, crude antibody and complete antibody,IgM. Rh blood group system and clinic work Transfusion and pregnacy [Clinic meaning]

Quantification of Blood Volume

Blood volume is an important determinant of systemic arterial pressure. Circulatory system is essentially a closed container including a volume of blood equal to approximately 5 liters or 70-80mL/Kg of the body weight (in kilograms).

4. Relation between blood volume and clinic

When you donate 10 % of total blood volume, your body compensates so that blood pressure does not change, and the volume is replaced through the normal ingestion of fluids. Volume loss up to 30-40 % of total blood volume can be tolerated if the loss is corrected within 30 min (e.g. artery contraction increases peripheral resistance but artery blood pressure can not maintain the normal levels which occur in symptoms such as light-headed, dazzled, force-lacked, etc) Blood loss more than 40 % of total blood volume will threaten the life, results in shock and the measures in the hospital should be immediately taken for life survival [Transfusion].

5. Principle of Transfusion
Transfusion is widely used in clinic treatment. Principle of transfusion*: 1. Identification of blood group must be taken before transfusion. 2. Cross-match test must be done before transfusion. 3. The same tpyes of blood group for transfusion should be firstly considered. 4. The different tpyes of blood group for transfusion should be very careful, small amount and slow import and if condition is better, changes in the same tpyes of blood group for transfusion.

Cross-match test for transfusion

RBC Donator RBC

Receiver

Serum
Main side of agglutination

Serum

Subordinary side of agglutination

Decision

+ -

+, +

Perfect match, transfusion No match, transfusion Transfusion under emergency

+: Agglutination; -: No agglutination

Types of Transfusion

According to source of transfusion, allogenetic transfusion (more use), autologous transfusion. According to component of transfusion, whole blood transfusion, transfusion of blood components Autologous transfusion has some advantages:
It decreases infection. It blocks syndrome (fever, hemolysis) induced by allogenetic transfusion. It stimulates bone marrow hemopoiesis towards RBC.

Transfusion of blood components is good.

Summarization
PLEASE TAKE DOWN

Consideration after class

1. Please describe classification and main effects of leucocyte. 2. What is the elementary process of blood

coagulation and main factors which have

participated in blood coagulation? 3. Please describe the principle of classification

and blood transfusion of ABO blood group

system.

Guide of Reference
. . . : , 2000. , , . . : , 2000. , , . . : , 2001. . . , : , 2005. , , . . : , 2000. , . . : , 1999. Ding L, Lu S, Batchu R, et al. Bone marrow stromal cells as a vehicle for gene transfer. Gene Ther, 1999, 6(9): 1611-1616. 8. Humeau L, Bardin F, Maroc C, et al. Phenotypic, molecular, and functional characterization of human peripheral blood, CD34+/Thy1+ cells. Blood, 1996, 87(3): 949-955. 9. Kaushansky K. Thrombopoietin: accumulating evidence for an important biological effect on the hematopoietic stem cell. Ann N Y Acad Sci, 2003, 996: 39-43. 10. Berne RM, Levy MN, Koeppen BMI, Stanton BA. Physiology, 5th ed, St Louis: Mosby Electronic Production, 2004. 11. Guyton AC, Hall JE. TEXTBOOK OF MEDICAL PHYSIOLOGY, 10th ed, Philadelphia: W.B. Saunders Co, 2000. 12. Berardi AC, Wang A, Levine JD, et al. Functional isolation and characterization of human hematopoietic stem cells. Science, 1995, 267(5194): 104-108. 13. Fox SI. Human physiology, 7th ed, New York: McGraw-Hill Co Inc, 2002. 14.Davies A, Blakeley AGH, Kidd C. Human physiology. Edinburgh: Churchill Livingston, 2001. 1. 2. 3. 4. 5. 6. 7.

Navigation for Web Address

1.http://bioresearch.ac.uk/browse/mesh/detai l/c0005811L0005811.html 2.http://www.inform.umd.edu/EdRes/Colleges /HONR/HONR269U/Jenn/ 3.http://www.ohsu.edu/cliniweb/G9/G9.188.h tml 4.http://www.mednote.co.kr/PHYSIOLOGY%2 0BLUE.htm 5.http://www.fpnotebook.com/HEM38.htm

BLOOD PHYSIOLOGY QUESTIONS ANSWERS

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