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measured by a bathroom scale or weight machine. [69]

2.3 Physiological Measurement:

In physiological measurement, we measure the blood pressure and pulse rate of control and patients.

 

Blood pressure:

Blood pressure measurement is the non-invasive measurement of the pressure exerted by the circulating blood on the walls of the body's arteries. The non-invasive blood pressure is taken using a sphygmomanometer, a hand bulb pump, and a cuff. [70]

How blood pressure checked?

A soft arm cuff is wrapped around the upper arm. A hand bulb pumps air into the cuff, gently squeezing the arm and temporarily interrupting the flow of blood. The pressure gauge reaches a peak. Then the cuff is slowly deflated, letting blood flow again. As the cuff deflates and the pressure gauge gradually decreases, the return of the blood flow through the main artery in the arm can be heard using a stethoscope. The reading on the pressure gauge when the pulse is first heard is systolic pressure (the peak pressure as the heart contracts). The reading when the pulse can first no longer be heard is the diastolic pressure (the lowest pressure as the heart relaxes between beats).[71]

Pulse rate:

The pulse rate is the rate at which heartbeats. Your pulse is usually called heart rate, which is the number of times a heart beats each minute (bpm). However, the rhythm and strength of the heartbeat can also be noted, as well as whether the blood vessel feels hard or soft. Changes in heart rate or rhythm, a weak pulse, or a hard blood vessel may be caused by heart disease or another problem.[72]

How pulse rate checked?

1. The first step to taking and checking for a normal rate is to locate the individual's wrist.

Then placing the first 3 fingers (pointer+middle+ring) just below the crease (where hand and wrist meet just under the thumb) press until can feel the radial pulse. Counting pulse rate is a simple way to find out how fast the heart is beating. [73]

2.4. Blood sample collection:

Blood is typically drawn from a vein, usually from the inside of the elbow or the back of the hand. The site is cleaned with germ-killing medicine (antiseptic). The health care provider wraps an elastic band around the upper arm to apply pressure to the area and make the vein swell with blood.                                           

Next, the health care provider gently inserts a needle into the vein. The blood collects into an airtight vial or tube attached to the needle. The elastic band is removed from their arm.                                                                                  

 Once the blood has been collected, the needle is removed, and the puncture site is covered to stop any bleeding.[74]

2.5 Procedure:

2.5.1Hematological analyzer:

1.      A complete blood count test were carried on Hematological analyzer (Medononic 620).This instrument is a fully automated hematology analyzer designed to measure up to 20 parameters using whole blood from an open inlet, closed tubes, micro pipettes 20 microlitre or pre-diluted blood.2.      The instrument employs the electronic impedance principle for cell counting and sizing and colorimetric method for measuring hemoglobin. A microprocessor is used to measure the parameters and to size the cells. During the count the processor checking the analyzing process for any irregularities. Size distributions are printed for all populations (RBC, PLT, and WBC).3.      The instrument has as standard a parallel and a serial output, which is user-programmable. For a built-in selectable program, the user can choose between different print formats.4.      The instrument is fully automatic, which means that the system is always powered on and performs automatically to perform check-and cleaning cycle to minimize user maintenance. [75] 2.5.2 Measuring principle:1.    RBC, WBC, and PLT concentration detection:·         Detection is accomplished using the electronic impedance principle and occurs in the orifice of the transducer.·        

The blood is diluted to 1:400(WBC & HBG) and 1:40000(RBC & PLT) through a precise shear system.The shear valve “cut” a very reproducible volume(25microlitre)from the aspired blood and dilute with an equal precise volume of diluents (or lyzer) to achieve the final dilution rate.·         Two separate measuring chambers and transducer are used, one for RBC/PLT and one for WBC/HGB analysis. This excludes any possibility of cross-contamination between the lyzer and the RBC/PLT dilution.·         A pressure is applied on top of the diluted sample and the diluted sample is pressed through an orifice (operature) of 80 micrometer diameter .Each side of orifice is equipped with a platinum electrode current and an electrical current is applied between  the electrode.·         When a cell is drawn into a constant current, flowing from the electrode through the orifice to a second electrode, the electrical conductivity is change s. This generates an equivalent voltage pulse.·         The amplitude of the pulse is directly proportional to the volume of the represented cell. The number of pulses corresponds to the number of cells detected.·         The PLT, RBC, and WBC parameters are measured on a precise aliquot of the sample. The amount of sample measured is determined by the volume of a precise glass column; called a metering tube. Two optical detectors are used to start and stop detection.·         The start detector is activated by the flow of the isotonic diluent through the metering tube. As the meniscus passes the optical path it causes a voltage change that activates the count and sizing circuitry of the system.·         As the isotonic diluent continues to flow upward into the metering tube it passes the stop detector .This action stop the count an analyzing process and the parameters and distribution curves are displayed. Due to this principle, the CA instruments perform absolute counts related to fixed volumes.·        

The instrument used a lower discriminator level at approx. 2.5 fl.All cells over this level are analyzed and counts are stored.·         The reproducibility is directly related to the total number of cells entering the orifice. The higher the concentration, the better the reproducibility. The instrument has a dilution ratio for RBC of 1:40,000 and the CV will, therefore, be less than 1% for the sample with an RBC number within the normal range. [75]

Normal ranges:·         RBC’s = 4.5-5.9million/cmm (Male)                      

=3.8-5.8 million/cmm (Female)·        

WBC’s = 4000-11000/cmm   (Adult)                        =6000-18000/cmm     (Children)·        

Platelets=1, 50,000-4, 50,000/cmm2.

Sizing RBC, WBC, and PLT:·        

The sizing is done in a matrix with the volume on the horizontal (x) axis and the number of the cells on the vertical(y) axis. [75]3.

WBC differentials:·        

The instrument utilizes a so-called mathematical differential, where the curves are analyzed within the software and 3 separate curves are built through a curve fitting method.·        

Hence, the software of the instrument is building an artificial matching distribution around the main populations. To do so, after the analyzing process, the instrument finds two main modes (=peaks) within the total distribution. Then matching of the 2 main populations takes place including extrapolation to the baseline. The remaining area that was not covered by the 2 main population is now classified as being the MID cell area which mainly consists of the Monocyte.

A third population is now calculated, representing this area. [73]

Normal ranges:·         Neutrophiles = 50-70%·        

Eosinophils = 01-06%·       

Basophils     = 00-01%·        

Lymphocytes = 25-45%·        

Monocytes     = 02-10%5.   

MCV (Mean Cell Volume RBCs):·        

The MCV parameter is derived from the RBC’s distribution curve. As the distribution curve has a maximum volume range of 250fl, the maximum channel also contains clumps of cells that are large then this volume. Therefore this channel is excluded from the MCV calculation. The MCV calculated from the volume position of the discriminating to 249fl.·        

In general, RBC count that is lower than 0.60 (displayed value) does not give an MCV/HCT value due to low statistical significance. [75]        

Normal ranges:·               MCV =             76-96 fl   

6. HGB (Hemoglobin Concentration):·        

The hemoglobin is determined from the same dilution as the WBC. For each sample a blank is measured as a reference; this means that any drift in reagent and cuvette-absorption or lamp is eliminated. The photometer system consists of a tungsten lamp, a cuvette with a length of 15 mm and a filter at a wavelength of 535 nm (bandwidth 20nm). [75]

Normal ranges:·         Hemoglobin =13-18g/dl         (Male)                                

=11.5-16.5g/dl   (Female)

7. MCH (Mean Cell Hemoglobin):·        

The MCH is a calculated value and is defined as HGB/RBC giving the mean HG B concentration in each red cell. [75] 

Normal range:·         MCH =             27-32 pg       

8. MCHC (Mean Cell Hemoglobin Concentration)·       

 

  The MCHC is a calculated value and it is defined as HGB/HCT.·         The MCHC is calculated from 3 measured parameters and therefore an excellent instrument stability check. MCHC =HGB/HCT HGB/ (MCV X RBC). [75]Normal range:·         MCHC =          30-35 g/dl

RESULT & DISCUSSION

 

Statistical Comparison Of RBC’s & Hb In Control & cancer Patients.

 

Parameters:

 

Control Samples  (n=40)

Cancer Patient Samples (n=40)

t-Test

p-Value

Mean+SD

Mean+SD

RBC’s (million/cmm)

4.5+0.4

4.4+0.7

1.06

0.3

Hb (g/dl)

12.6+1.1

11.0+2.3

1.94

0.1

                                                                                                Table 3.1, shows the mean values, of RBCs and Hb of normal and cancer patients. Mean RBCs of normal and cancer patients were 4.5+0.5 and 4.4+0.7 respectively, t-test value is 0.11 and for all probability of RBC’s is0.9. This indicates that the RBCs have no significant difference in their levels. Some cancer treatments, especially chemotherapy and radiation therapy, may cause a decrease in your body’s RBCs. This condition is known as anemia. Blood loss, either from surgery or from specific cancers, can also cause or worsen anemia. [76]

 

 The mean Hb of normal and cancer patients was 12.6+1.1 and 11.08+1.1; the t-test value is 1.9 and the p-value of Hb is 0.1. This indicates that Hb has a significant difference in their level which is slightly low in cancer patients. Several experimental studies have shown that low levels of hemoglobin (Hb) correlate with diminished partial oxygen pressure (pO2) in the tumor. [77- 80]

 Many studies have shown that the use of erythropoietin (EPO) in oncologic patients increases Hb levels. [81-86].

Table.3.2

Statistical Comparison Of Absolute Indices In Control & cancer Patient.

 

Parameters:

 

 

 

Control Samples (n=40)

 

Cancer Patient Samples (n=40)

 

t-Test

p-Value

Mean+SD

Mean+SD

MCV (fl)

81.7+6.7

78.1+9.7

0.15

0.9

MCH (pg)

27.9+5.6

25.6+3.5

0.4

0.7

MCHC (g/dl)

35.0+2.3

32.7+0.8

0.1

0.9

 

Table-3.2 shows the mean value of MCV, MCH, and MCHC of normal and cancer patients. The mean value of MCV for normal and cancer patients was 81.7+6.7 and 78.1+9.7 and the t-test is 0.5 and for all probability of MCV is 0.6.which shows that the MCV value of cancer patients is slightly low than normal.

 And the mean value of MCH for normal and cancer patient was 27.9+5.6 and 25.6+3.5. And the t-value is 0.4 and the p-value of MCH is 0.7.this indicates that MCH is also low in cancer patient then normal.

While mean value of MCHC for a normal person and cancer patient was 35.0+2.3 and 32.7+0.8.While t-test value is 0.1 and the p-value of MCHC is 0.9.It is also low in cancer patients.                                                                                                            

                                             

 

 

 

Table: 3.3

Statistical Comparison of WBC’s In Control & cancer Patient.

 

Parameters:

 

 

 

Control Samples (n=40)

Cancer Patient Samples (n=40)

t-test

p-Value

Mean+SD

Mean+SD

TLC (cmm)

9490+4279.4

8332.5+3644.9

0.78

0.5

Neutrophils (%)

70.6+11.69

52.8+26.3

0.26

0.8

Lymphocytes (%)

26.6+11.3

23.6+13.7

0.97

0.4

Eosinophils (%)

1.07+0.6

2.7+0.7

10.35

0.001

Monocytes (%)

1.7+0.8

2.9+0.7

6.75

0.001

Basophils (%)

00+00

1.4+0.5

16.37

0.001

                                                                                                                    

Table-3.3, shows the mean values, t-values and p-values of TLC, Neutrophil, Lymphocytes, Eosinophils, Monocytes and Basophils of normal persons and cancer patients. The mean value of TLC for normal persons and cancer patients was 9490+4279.4 and 8332.5+3644.9, the t-value is 0.78 and the p-value of TLC is 0.5.which indicates that TLC level is low in the cancer patient.

The mean value of neutrophils for normal and cancer patients was 70.6+11.6 and 52.8+26.3,t-value is 0.26 and p-value of neutrophils was 0.8. It also shows that the neutrophil is low in cancer patients. These cancers and cancer treatments may cause a condition called neutropenia, an abnormally low level of neutrophils. [76]

The mean value of lymphocytes for normal and cancer patients is 26.6+11.3 and 23.6+13.7, and t-test

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