ISSN NO 2320-5407

International Journal of Advanced Research (2013), Volume 1, Issue 5, 71-78

Journal homepage: http://www.journalijar.com

INTERNATIONAL JOURNAL
OF ADVANCED RESEARCH

RESEARCH ARTICLE
Evaluation of Anticancer properties of Taxusbaccata and Badri cow urine in mice: Clinicohematological
study
Ankita Joshi, and R.S. Chauha
1. Post Doctoral Fellow, Cell Culture Laboratory, Institute of Biotechnology, G.B. Pant University of Agriculture
and Technology, Patwadangar-263128, Nainital, Uttarakhand, INDIA.
2. Campus Director, Institute of Biotechnology, G.B. Pant University of Agriculture and Technology, Patwadangar263128, Nainital, Uttarakhand, INDIA.

Manuscript Info

Abstract

Manuscript History:

In the present investigation, the anticancerous effect of Taxusbacaata and

distilled Badri cow urine was studied in mice for clinicohematology and
body weight for a period of six month at an interval of 15 days. The study
revealed that the values of total leucocyte count (TLC), absolute lymphocyte
count (ALC) and absolute neutrophil count (ANC) were significantly
increased in the treated groups of mice either by CUD alone and in
combination withTaxusbaccata extracts. At180th day, it was found that there
was an increase in body weight, Hemoglobin content (Hb), total erythrocyte
count (TEC), total leucocyte count (TLC), absolute lymphocyte count (ALC)
and absolute neutrophil count (ANC) levels in CUD +A treated group as
23%, 23.99%, 41%, 40%, 40.31%, and 40.13%, respectively. This clearly
indicate the, increase in vitality and defence mechanism of body which in
turn helps in further healing of cancer.

Received: 12 June 2013

Final Accepted: 23 June 2013
Published Online: July 2013

Key words:
Badricow urine,
Taxusbaccata, Mice,
Body weight, hematology.

Copy Right, IJAR, 2013,. All rights reserved.

Introduction
Cancer is a disease involving dynamic changes in
genome and is characterized by the uncontrolled,
uncoordinated and purposeless proliferation of
malignant cells and their ability to spread, either by
growth in the adjacent tissue through invasion or by
implantation at distant sites through metastasis.
World cancer report issued by International Agency
for Research on Cancer (IARC) reported in 2003 that
cancer rate is set to increase at an alarming rate
globally. The 5-year relative survival rate for all
cancers diagnosed between 1999-2005 is 68%, up
from 50% in 1975-1977. (Kinzler and Vogelstein,
2002)(Jemal et al., 2011).
In India about 70% of population obtains medical
help from private practitioners and half of those who
seek medicinal help obtain it from alternative and
traditional medicine (Kumar et al., 2004). Poverty
and socioeconomic status are other hurdles in
treatment (Pal and Mittal, 2004). American cancer
society defines complementary and alternative
medicines (CAM) simply as anything which is not

conventional (Zollman and Vickers, 1999; Park et al.,

2003). There are various CAM used for cancer
patient worldwide viz. Herbal medicine, acupuncture,
Ayurveda, biological agents, traditional Chinese
medicines, meditation and yoga etc. However, use of
herbs for cancer treatment is very popular throughout
the world.
Distilled cow urine protects DNA and
repairs it rapidly as observed after damage due to
pesticides. It protects chromosomal aberrations by
mitocycin in human leukocyte. Cow urine helps the
lymphocytes to survive and not to commit suicide
(apoptosis). Pathogenic effect of free radicals are
prevented through cow urine therapy. Use of cow
urine one can get the charm of a youth as it prevents
the free radicals formation. Taxus baccata,
commonly known as THUNER, which is mainly
found in high altitude area like, Patwadangar,
Nainital India also had anticancer and antiviral
properties. It is a small to medium-sized evergreen
tree, growing 10-20 m tall, exceptionally up to 28 m.
It is relatively slow growing, but can be very longlived, with the maximum recorded trunk diameter of

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International Journal of Advanced Research (2013), Volume 1, Issue 5, 71-78

4 m probably only being reached in around 2,0004,000 years. Thuner is the oldest plant at high altitude
region of Uttarakhand. Most parts of the tree are
toxic, except the bright aril surrounding the seed,
enabling ingestion and dispersal by birds. The major
toxin is the alkaloid taxane. Phytochemical analysis
of extracts of leaves and bark showed the presence of
lignans, flavonoid, glycosides, sterols, sugar, amino
acid, and triterpenoid, alkaloids, steroids, tannins,
mucilage, fixed oil, phenolic compounds and
protein.The leaves are the principal source of taxol;
the anti-cancer drug, but has not been widely
exploited in this connection (Hartzell, 2003).
Considering the severity of cancer as a
disease of man and animals and complexity of
therapeutic approaches and their harmful side effects,
it was planned to study the effect of Taxusbaccata
preparation along with cow urine distillate in mice as
measured through clinical haematology

Materials and Methods

1.

Extract preparation

Extracts of leaves and bark of T. baccata were

prepared by applying the standard methods with
different solvents like; Aqueous, ethanol, methanol
and ether as described by Govindachariet al., (1999)
and Udupaet al., (1995).
In-vivo study
2. Experimental design
Present study was performed in mice maintained in
the experimental animal house in Institute of
Biotechnology, G.B. Pant University of Agriculture
and Technology, Patwadangar, Nainital, Uttarakhand,
India. A total of 97 animals were equally divided into
11 groups. The mice were housed in clean
polypropylene cages and fed adlibitum with
commercially available feed and water. The
experiment was carried out in accordance with the
Institutional Animal Ethical Committee (IAEC), G.B.
Pant University of Agriculture and Technology,
Pantnagar, Uttarakhand, India. The 11 groups
were:Control group(9 mice), Negativecontrol(DEN
treated mice)(8 mice), CUD(without DEN)(8 mice),
A (Aqueous extract of leaves of Taxusbaccata)(8
mice),
(Ethanolic
extract
of
leaves
of
Taxusbaccata)(8 mice), G(Methanolic extract of bark
of Taxusbaccata)(8 mice), H(Ether extract of Bark of
Taxusbaccata)(8
mice),
CUD(Cow
Urine
Distillate)(with DEN)(8 mice), CUD+A(8 mice),
CUD+B(8 mice), CUD + G(8 mice), CUD+H (8
mice).
Single dose of diethyl nitrosamine (DEN) @ 200
l/kg body weight was given to each mice of
negative control group and tests groups. 500 ml of

each extract were made by adding 20% of extract in

500ml of distilled water (Kumar et al., 2004b). The
mice of 9 test groups were given different extracts of
taxusbaccata alone and in combination with CUD
(2ml/day/mice), daily p.o., from day 1 for 6 months;
however, the mice of negative control group were
maintained with routine feed and water.Body weight
of mice were taken regularly at an interval of 15 day
till the end of the experiment.Total leucocyte count
(TLC), absolute neutrophil count (ANC), absolute
leucocyte count (ALC), hemoglobin, total erythrocyte
count (TEC) and hemoglobin (Hb) content of all the
experimental animals in different groups were
determined regularly at an interval of 15 day till the
end of the experiment as per in standard procedures
(Chauhan, 2005).

Results
In-vivo study was carried out in mice using DEN as
carcinogen and plant extracts alone and/or
combination with CUD as test material for a period
six month.
Body Weight
Body weight of mice were taken in gram at an
interval of 15 days till the end of experiment. Data of
body weight change during experiment were givens
in table-1. Initially, the mean bodyweight of control
was 21.471.21 gm and after 6 month the mean body
weight of mice was 25.861.87 gm. In DEN
(negative control) treated group the initial mean body
weight at zero day of experiment was 22.731.33 gm,
which decreased to 19.161.81 gm at the end of
experiment. But in CUD treated group mean body
weight at zero day was 22.431.36 gm and at the end
of experiment it was 23.911.21 gm.CUD treated
group in which the carcinogen has been given, the
initial mean body weight at zero day was 21.421.56
gm. After the end of experiment, the mean body
weight was 21.061.91 gm. In test group A, the zero
day mean body weight 23.131.54 gm, which
marginally increased at the end of experiment to
23.521.01gm.In the group CUD+A the mean in
body weight at zero day was 21.361.47 gm which
was 26.480.902 at the end of experiment. In the
group CUD+B, the mean body weight was
22.761.44 gm at zero day and was 24.801.09 gm at
the end of experiment. In CUD+G and CUD+H
groups, the mean body weight at zero day was
22.971.37 gm and 22.811.21 gm which was
increased to 24.670.941 gm and 24.601.01 gm at
the end of the experiment. In group B, G and H, the
mean body weight at zero day was found 22.811.26
gm, 22.411.32 gm and 22.511.28 gm respectively
and at the end of experiment the body weight reaches

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to

International Journal of Advanced Research (2013), Volume 1, Issue 5, 71-78

23.401.02, 23.331.91 and 23.281.89,

respectively.
Table 1: Body weight in gm of experimental mice at an interval of 15 days (MeanSE).

Groups/Days

0 Day

15

30

45

60

75

90

105

120

135

150

165

180

Control
CUD (no
DEN)
DEN

21.471.21*
22.431.36*

22.521.32
22.481.29*

22.921.47
22.511.33

23.171.07
22.541.42*

23.191.36
22.611.62**

24.311.41
22.791.46

24.561.53
22.941.53

25.071.43
23.081.67*

25.190.947
23.481.82

25.371.12
23.541.29*

25.691.49
23.671.68

25.720.989
23.821.92

25.861.87
23.911.21**

22.731.33**

23.121.42

23.491.36*

23.791.21*

24.011.36

24.671.17**

24.921.07*

25.160.941

24.011.21*

23.721.06

22.091.09*

21.871.31*

19.161.81

CUD

21.421.56*

21.621.41*

21.711.52*

22.071.02

22.671.17**

22.891.19

23.121.23*

22.921.16

22.660.941

22.521.12

22.261.07

22.121.13*

21.061.91*

23.131.54*

23.631.71

24.761.23

24.931.32**

25.111.16*

25.721.21

25.911.47

24.371.41**

24.871.31

23.851.03**

23.711.07

23.681.13

23.521.01*

22.811.26

23.121.21**

23.711.28

23.911.17

24.111.24*

24.521.31**

25.031.28*

24.761.19*

24.101.07*

23.811.23

23.711.16*

23.671.09

23.401.02*

22.411.32

22.911.61*

23.141.10**

23.571.17

23.871.36*

24.101.30

24.471.28

24.321.21*

24.101.07

23.731.22*

23.640.940

23.501.11

23.331.91*

22.511.28*

22.681.31

22.911.12*

23.411.21**

23.621.31

23.821.40*

23.981.33

23.901.21

23.811.17

23.631.07

23.481.21

23.391.07**

23.281.89

A+CUD

21.361.47**

21.731.32*

22.071.12**

22.391.39

22.871.42*

23.011.25

23.931.61**

24.031.71*

24.180.981**

25.811.42*

25.911.07*

26.361.03

26.480.902**

B+CUD

22.761.44*

22.911.51*

23.571.07

23.911.40**

23.961.31

24.031.61

24.181.42*

24.431.27

24.891.19

24.971.36*

24.911.17

24.881.13

24.801.09

G+CUD

22.971.37

23.071.27

23.461.21*

23.711.41**

23.961.31*

24.101.31*

24.571.19

24.811.17**

24.951.16*

24.891.27*

24.841.11

24.721.08*

24.670.941

H+CUD

22.811.21*

22.901.20*

22.961.17

23.071.31

23.481.32**

23.691.20

23.841.23

23.961.19

24.971.23

24.901.21

24.781.31**

24.671.08

24.601.01*

Significant difference in comparison to control (*p0.5 and **p0.01)

Table 2: Total erythrocyte count (TEC) (x 106/cumm) of experimental mice at an interval of 15 days (MeanSE).
Groups/Days

15

30

45

60

75

90

105

120

135

150

165

180

Control

6.820.13

6.860.17

6.920.27

6.970.16

7.060.31

7.140.24

7.160.41

7.180.37

7.190.34

7.200.17

7.210.19

7.230.23

7.250.27

CUD (NO
DEN)

6.290.52*

6.360.41*

6.520.63

6.700.13*

6.830.46

6.940.32

7.030.21**

7.190.61*

7.220.73*

7.250.49*

7.290.36

7.340.51**

7.400.42

DEN

6.870.51

6.890.62**

6.960.71*

6.980.18*

7.080.42*

7.110.52*

7.010.63

6.940.87*

6.660.33*

6.060.61

5.040.12**

4.710.43

3.610.46*

CUD

6.420.83

6.480.17

6.630.21*

6.690.47*

6.780.82*

6.830.27*

6.910.16*

7.030.51*

7.140.21

7.010.32**

6.810.27

6.710.61

6.680.53

6.310.37**

6.380.41

6.480.28*

6.630.46*

6.890.72*

6.960.69

7.090.33*

7.470.38

7.340.28

7.120.48

7.190.59*

7.230.72

7.320.58

6.330.61

6.450.42*

6.610.47

6.750.51

6.950.80

7.080.72**

7.220.61

7.450.56

7.290.51*

7.090.62

7.150.68*

7.210.52*

7.290.57**

6.390.41

6.410.39

6.540.41*

6.710.43

6.900.53

7.050.62**

7.170.47

7.390.54**

7.060.57

6.890.62

7.050.41*

7.150.46

7.200.37

6.370.52*

6.390.59*

6.530.47

6.680.45

6.700.52**

6.720.58

6.960.43

7.150.48

7.410.51**

7.690.48

7.350.61

7.230.42*

7.180.41**

A+CUD

6.310.97**

6.460.818

6.790.17*

6.980.42**

7.090.78

7.350.18*

7.490.19**

7.830.63*

7.980.76

8.380.84

8.730.39**

8.830.36*

8.900.47

B+CUD

6.370.53

6.400.61**

6.660.59

6.770.41*

6.810.47*

6.960.53

7.090.12

7.180.35

7.210.61*

7.400.69**

7.770.63

8.150.71

8.590.62*

G+CUD

6.420.59*

6.440.53

6.790.49

6.830.50

6.930.60

7.070.59*

7.180.57

7.310.51

7.770.53

7.860.59

7.720.47*

8.190.44

8.510.42

H+CUD

6.350.61

6.400.57

6.550.64**

6.720.51*

7.130.57

7.450.63

7.680.53*

7.880.56*

8.040.47**

8.130.52

8.290.56

8.440.48**

8.480.43*

Significant difference in comparison to control (*p0.5 and **p0.01)

Haematological parameters
Data of TEC is expressed in number of
cellsx106/cumm and is mentioned in Table 2. Initially
the TEC of control was 6.820.13 x 106/cumm and
after 6 month, TEC of experimental mice was
7.250.27 x 106/cumm. In DEN treated (negative
control) group the initial TEC at zero day of
experiments was 6.870.51 x 106/cumm which

decreased to 3.610.46 x 106/cumm significantly at

the end of experiment. But in CUD treated group, the
TEC at zero day was 6.290.52 x 10 6/cumm and
7.400.42 x 106/cumm at the end of experiment.
CUD treated group in which the carcinogen had also
been given, the initial TEC at zero day was 6.420.83
x 106/cumm. After the end of experiment, it was
6.680.53x 106/cumm. In test group A, the zero day

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TEC was 6.310.37 x 106/cumm. The TEC decreased

to 7.320.58 x 106/cumm at 180 day of experiment.
In group CUD+A the total erythrocyte count, at zero
day was 6.310.97 x 106/cumm which was increased
to 8.900.47 x 106/cumm at the end of experiment.
Group CUD+B had 6.370.53 x 106/cumm TEC at
zero day and 8.590.62 x 106/cumm at the end of
experiment, respectively. In CUD+G and CUD+H
groups the TEC at zero day were 6.420.59 x
106/cumm and 6.350.61 x 106/cumm, respectively
and were 8.510.42 x 106/cumm and 8.480.43 x
106/cumm at the end of the experiment. In group B, G
and H, the TEC at zero day was found 6.330.61 x
106/cumm, 6.390.41 x 106/cumm and 6.370.52 x
106/cumm, respectively and at the end of experiment
increased to 7.290.57 x 106/cumm, 7.200.37 x
106/cumm and 7.180.41 x 106/cumm.
Data of TLC is expressed in no. of cells x 103/cumm
and is presented in Table-3.Initially the TLC count of
control group was 8.910.30 x 10 3/cumm and after 6
month the TLC was 12.110.04 x 103/cumm. In DEN
(negative control) group the initial TLC at zero day
of experiments was 9.123.20 x 103/cumm which
was decreased to 3.021.45 x 103/cumm. But in CUD
treated group, the TLC at zero day was 8.654.31 x
103/cumm and 9.534.67 x 103/cumm at the end of
experiment.
In CUD treated group, the initial
TLC at zero day was 9.025.31 x 103/cumm which
decreased to 7.813.11 x 103/cumm. In test group
like A, the zero day TLC was 9.085.68 x 10 3/cumm
which
was
decreased
to
8.643.08
x
103/cumm.CUD+A had the zero day mean TLC
count as 8.974.02 x 103/cumm which was increased
to 12.612.17 x 103/cumm, at the end of experiment.
Group CUD+B has 8.896.31 x 103/cumm at zero
day and was 10.553.18 x 103/cumm at the end of
experiment.In group B, G and H, the TLC at zero day
was observed as 9.016.81 x 103/cumm, 9.106.07 x
103/cumm and 9.077.03 x 103/cumm, respectively
and at the end of experiment 8.593.19 x 10 3/cumm,
8.333.61 x 103/cumm and 8.304.1 x 103/cumm
respectively. In groups CUD+G and CUD+H the
TLC at zero day was 9.115.98 x 103/cumm,
9.036.19 x 103/cumm and was 10.213.81 x
103/cumm and 10.053.14 x 103/cumm at the end of
the experiment, respectively.
Data of ALC is expressed in no. of cells x
103/cumm and is presented in Table-4.Initially the
ALC count of control group was 4.300.69 x
103/cumm and after 6 month the ALC was 5.920.98
x 103/cumm. In DEN (negative control) group the
initial ALC at zero day of experiments was 4.410.81

x 103/cumm which was decreased to 1.360.47 x

103/cumm. But in CUD treated group, the ALC at
zero day was 4.130.70 x 103/cumm and 4.630.43 x
103/cumm at the end of experiment.In CUD treated
group, the initial ALC at zero day was 4.430.84 x
103/cumm which decreased to 3.760.53 x
103/cumm. In test group like A, the zero day ALC
was 4.420.91 x 103/cumm which was decreased to
4.190.27 x 103/cumm.CUD+A had the zero day
mean ALC count as 4.390.87 x 103/cumm which
was increased to 6.160.42 x 103/cumm, at the end of
experiment. Group CUD+B has 4.360.89 x
103/cumm at zero day and was 5.160.80 x
103/cumm at the end of experiment.In group B, G and
H, the ALC at zero day was observed as 4.400.94 x
103/cumm, 4.480.73 x 103/cumm and 4.440.84 x
103/cumm, respectively and at the end of experiment
4.180.94 x 103/cumm, 4.010.37 x 103/cumm and
3.980.77 x 103/cumm respectively. In groups
CUD+G and CUD+H the ALC at zero day was
4.410.80 x 103/cumm, 4.420.79 x 103/cumm and
was 4.900.28 x 103/cumm and 4.890.11 x
103/cumm at the end of the experiment, respectively.
Data of ANC is expressed in no. of cells x
103/cumm and is presented in Table-5.Initially the
ANC count of control group was 4.570.72 x
103/cumm and after 6 month the ANC was 6.000.99
x 103/cumm. In DEN (negative control) group the
initial ANC at zero day of experiments was
4.690.83 x 103/cumm which was decreased to
1.490.52 x 103/cumm. But in CUD treated group,
the ANC at zero day was 4.380.78 x 10 3/cumm and
4.710.47 x 103/cumm at the end of experiment.
In CUD treated group, the initial ANC at
zero day was 4.510.97 x 103/cumm which decreased
to 3.880.56 x 103/cumm. In test group like A, the
zero day ANC was 4.600.97 x 103/cumm which was
decreased to 4.300.31 x 103/cumm.CUD+A had the
zero day mean ANC count as 4.460.89 x 10 3/cumm
which was increased to 6.250.45 x 10 3/cumm, at the
end of experiment. Group CUD+B has 4.410.92 x
103/cumm at zero day and was 5.240.82 x
103/cumm at the end of experiment.In group B, G and
H, the ANC at zero day was observed as 4.580.99 x
103/cumm, 4.560.75 x 103/cumm and 4.510.87 x
103/cumm, respectively and at the end of experiment
4.260.96 x 103/cumm, 4.110.38 x 103/cumm and
4.090.79 x 103/cumm respectively. In groups
CUD+G and CUD+H the ANC at zero day was
4.520.84 x 103/cumm, 4.500.85 x 103/cumm and
was 5.010.29 x 103/cumm and 4.960.12 x
103/cumm at the end of the experiment, respectively.

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Table 3: Total leucocyte count (TLC) (x 103/cumm) count of experimental mice at an interval of 15 days (MeanSE).
Groups/Days

15

30

45

60

75

90

105

120

135

150

165

180

Control

8.910.30

9.060.17

9.380.32

9.910.37

10.080.51

10.210.62

10.530.02

10.730.12

11.050.81

11.320.91

11.510.07

11.870.47

12.110.04

CUD (NO
DEN)
DEN

8.654.31

8.714.36**

8.864.51*

8.934.70

9.085.01*

9.135.21

9.183.24*

9.204.22

9.274.56**

9.324.61

9.384.28

9.464.39*

9.534.67

9.123.20**

9.213.52*

9.434.07**

9.893.81

8.073.61

7.423.82

6.412.01

6.123.10*

5.872.12*

4.313.19

4.162.16

4.081.87*

3.021.45*

CUD

9.025.31*

9.135.27

9.315.17

9.674.81**

9.894.72

10.074.55

9.914.31**

9.734.24

9.414.27*

9.013.69

8.233.16**

8.113.08*

7.813.11

9.085.68*

9.155.51*

9.285.42

9.575.23

9.795.05**

9.984.93

9.784.61*

9.614.34

9.324.21*

8.914.07

8.833.21

8.793.12

8.643.08

9.016.81*

9.116.72

9.196.61**

9.416.52

9.616.37*

9.775.83*

9.585.61

8.915.37

8.875.21*

8.815.08

8.794.61*

8.684.52

8.593.19**

9.106.07

9.165.91*

9.245.82

9.335.74**

9.515.41*

9.675.34*

9.485.28

8.875.05

8.764.81

8.694.23*

8.594.07

8.413.82

8.333.61

9.077.03

9.116.87

9.156.67**

9.216.41

9.385.91

9.475.80

9.315.47*

8.835.32**

8.735.16

8.624.77*

8.574.41*

8.464.21*

8.304.10

A+CUD

8.974.02**

9.214.47

9.493.32

9.893.77*

10.082.01*

10.332.61*

10.693.16*

10.934.03

11.312.12**

11.673.41*

11.914.91*

12.253.53*

12.612.17**

B+CUD

8.896.31*

9.126.27*

9.256.08

9.495.61*

9.685.32*

9.875.17*

9.675.03

9.474.71

9.734.57

9.844.31

10.084.17

10.674.08*

10.553.18

G+CUD

9.115.98

9.185.81*

9.275.61

9.395.47*

9.595.27

9.715.17

9.515.09**

9.304.81

9.264.67

9.424.51

10.024.41**

10.174.32*

10.213.81*

H+CUD

9.036.19

9.136.01

9.195.81**

9.275.72

9.445.51

9.595.42*

9.405.31

9.315.06*

9.124.87*

9.094.62*

9.514.41

9.884.17

10.053.14

Significant difference in comparison to control(*p0.5 and **p0.01)

Table 4: Absolute lymphocyte count (ALC) (x 106/cumm) of experimental mice at an interval of 15 days (MeanSE).
Groups/Days

15

30

45

60

75

90

105

120

135

150

165

180

Control

4.300.69

4.410.42

4.590.24

4.810.49

4.920.19

4.980.33

5.170.54

5.230.68

5.410.57

5.530.28

5.620.31

5.810.73

5.920.98

CUD (NO
DEN)
DEN

4.130.70*

4.240.72

4.310.69*

4.370.58*

4.430.42

4.450.39*

4.480.69**

4.490.84

4.520.92*

4.540.14

4.550.28*

4.610.63*

4.630.43

4.410.81*

4.430.61

4.560.71

4.860.24*

3.920.92**

3.590.43*

3.050.64

2.940.59

2.810.36

2.080.31*

1.920.27*

1.900.17

1.360.47*

CUD

4.430.84*

4.410.64

4.540.84

4.740.18

4.830.29

4.850.36

4.850.47*

4.720.53*

4.590.61

4.350.74*

3.960.89

3.890.92

3.760.53

4.420.91

4.440.738

4.510.42**

4.670.75

4.780.94*

4.860.86

4.780.73

4.660.21**

4.530.28*

4.360.69

4.280.74**

4.250.18**

4.190.27*

4.400.94

4.420.57

4.440.55

4.600.85

4.690.69

4.770.54**

4.650.22*

4.330.51

4.340.49*

4.260.16

4.240.21*

4.220.86

4.180.94*

4.480.73*

4.410.82**

4.510.98*

4.560.37

4.640.76

4.760.41

4.620.47*

4.320.53

4.270.84

4.230.61

4.150.65

4.100.21*

4.010.37*

4.440.84

4.400.34*

4.480.76

4.500.26**

4.540.86*

4.630.16

4.560.36

4.300.61*

4.240.42*

4.150.69**

4.160.48

4.090.59*

3.980.77

A+CUD

4.390.87

4.490.29

4.620.59

4.840.43*

4.900.33

5.020.77

5.210.61

5.330.98

4.550.71

5.710.14

5.830.21*

5.980.63

6.160.42**

B+CUD

4.360.89**

4.470.19*

4.510.41**

4.620.82*

4.740.27

4.830.87*

4.720.91**

4.610.84**

4.730.16**

4.760.72

4.910.18

5.210.68*

5.160.80

G+CUD

4.410.80*

4.480.47

4.530.63

4.580.92

4.650.46**

4.740.18*

4.660.78

4.560.20

4.520.91

4.570.39*

4.870.65*

4.910.21

4.900.28

H+CUD

4.420.79

4.450.77

4.480.27*

4.520.39

4.610.65

4.690.23

4.590.88

4.540.49

4.460.64*

4.430.72*

4.630.47

4.860.30*

4.890.11*

Significant difference in comparison to control (*p0.5 and **p0.01)

Table 5: Absolute Neutrophil count (ANC) (x 106/cumm) of experimental mice at an interval of 15 days (MeanSE).
Groups/Days

15

30

45

60

75

90

105

120

135

150

165

180

Control

4.570.72

4.500.43

4.670.29

4.900.51

5.000.23

5.070.38

5.220.55

5.320.69

5.500.59

5.610.37

5.710.37

5.900.77

6.000.99

CUD (NO
DEN)
DEN

4.380.78**

4.330.74**

4.400.72

4.440.60

4.520.45*

4.530.43

4.560.72

4.570.86*

4.600.97

4.620.19

4.670.34

4.700.65

4.710.47*

4.690.83

4.560.65**

4.680.75

4.920.26

4.010.96

3.680.46

3.140.67**

3.000.61*

2.900.41

2.130.34

2.000.31

1.990.23*

1.490.52

CUD

4.510.87

4.500.69*

4.600.89*

4.810.21

4.920.31

4.990.37

4.930.49

4.830.57

4.670.66**

4.470.77

4.070.92**

3.990.95*

3.880.56**

4.600.97*

4.530.79

4.610.46

4.750.77

4.870.98

4.960.89*

4.860.74**

4.780.26

4.610.32

4.420.75*

4.390.77

4.360.23

4.300.31

4.580.99*

4.510.58**

4.560.57**

4.680.87*

4.780.69*

4.850.56

4.770.26

4.400.55

4.400.51

4.380.19*

4.350.26

4.310.89

4.260.96

4.560.75

4.540.86

4.620.99

4.640.39*

4.730.79*

4.800.43**

4.710.49

4.410.54**

4.350.85*

4.310.66*

4.260.66

4.180.24

4.110.38

4.510.87**

4.520.37

4.530.80*

4.580.29

4.660.88

4.710.17*

4.610.38

4.390.63

4.320.46

4.270.74

4.250.53*

4.200.61

4.090.79

A+CUD

4.460.89*

4.580.31*

4.700.61**

4.920.45*

4.990.35**

5.110.78*

5.320.64*

5.410.99*

5.620.74*

5.800.15**

5.910.23

6.100.65**

6.250.45*

B+CUD

4.410.92

4.510.23

4.630.46

4.730.86

4.800.29*

4.910.89

4.800.93

4.700.86

4.810.18

4.890.74*

5.000.20*

5.300.69

5.240.82

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G+CUD

4.520.84

4.560.49

4.610.66*

4.670.95*

4.770.48

4.800.20

4.730.79**

4.620.23

4.600.95*

4.680.44

4.980.67

5.000.24*

5.010.29*

H+CUD

4.500.85*

4.540.81*

4.560.29

4.610.40

4.700.66*

4.750.25

4.670.90

4.610.50*

4.530.65

4.510.73

4.720.49**

4.950.32

4.960.12

Table 6: Haemoglobin content (Hb) of experimental mice at regular interval of 15 days (gm%, meanSE).
Groups/Days

15

30

45

60

75

90

105

120

135

150

165

180

Control

12.451.42

12.471.51

12.531.52

12.511.42

12.551.47

12.501.44

12.521.41

12.551.37

12.561.35

12.581.39

12.621.34

12.661.21

12.681.36

CUD(NO
DEN)

12.191.31*

12.211.42

12.271.51*

12.491.36

12.621.47*

12.851.51

12.991.62*

13.411.73*

13.531.81*

13.661.74*

13.701.32

13.761.21*

13.781.19

DEN

12.401.36*

12.421.38

12.431.29

12.451.31**

12.461.33

12.251.36*

12.081.39

11.821.36*

11.651.30

10.521.32*

10.091.21

9.361.19*

8.021.07

CUD

12.321.31

12.451.28**

12.861.33

12.901.37

13.061.29*

13.401.42

13.871.47

13.671.40*

13.461.33**

13.221.28*

12.791.21*

12.091.19**

12.271.17**

12.431.27

12.841.29

12.871.32*

13.071.33

13.381.38*

13.851.41

14.041.43

13.951.47

13.801.27

13.761.23*

12.341.28
13.711.19

12.311.40

12.391.35*

12.791.25

12.831.38**

13.031.27

13.301.30

13.791.37**

14.021.39*

13.951.41*

13.751.26

13.711.19**

13.671.21**

13.561.18

12.341.28*

12.371.21

12.721.24**

12.791.41

13.041.32**

13.251.19**

13.721.29

13.961.26

13.911.25*

13.701.32*

13.651.27

13.611.21

13.491.29

12.341.42

12.361.40

12.681.23**

12.741.21

12.951.31*

13.101.22

13.671.28

13.871.27

13.821.30

13.651.33*

13.591.24**

13.551.32*

13.401.25

A+CUD

12.411.48*

12.481.47

12.891.36**

12.941.39

13.081.33*

13.421.41*

13.891.52*

14.061.47**

14.641.37

14.821.42

15.071.37

15.211.27

15.381.31*

B+CUD

12.291.42*

12.401.37

12.811.27

12.851.36**

13.051.31

13.351.32

13.811.39

14.011.41

14.221.42

14.481.25

14.741.21*

14.891.22*

14.901.17*

G+CUD

12.271.41

12.361.39**

12.761.21

12.801.29*

13.011.25

13.271.28

13.761.31*

13.981.30*

14.031.27*

14.331.21**

14.691.23

14.741.17

14.861.14

H+CUD

12.301.39*

12.351.35

12.701.27*

12.761.23

12.981.33**

13.121.29*

13.701.20

13.901.24

13.971.26

14.081.31

14.621.28*

14.701.22

14.781.11**

Significant difference in comparison to control (*p0.5 and **p0.01)

Data of Hemoglobin is expressed in gm% and is

presented in Table-6. Initially the Hb content of
control was 12.451.42gm% and after 6 month the
Hb content of mice was 12.681.36gm%. In DEN
(negative control) group the initial Hb content at zero
day of experiment was 12.401.36gm%, which
decreased to 8.021.07gm%, at the end of
experiment. But in CUD treated group, their Hb
content at zero day was 12.191.31gm% and
13.781.19gm% at the end of experiment. In CUD
treated group in which the carcinogen has been given,
the initial Hb content at zero day was
12.321.31gm%, after the end of experiment the Hb
content was 12.091.19gm%. In test group A, the
zero day Hb content was 12.271.17gm%. The Hb
content increased to 13.631.13gm% at the end of
experiment. In group B, G and H, the Hb content at
zero
day
was
found
12.311.40gm%,
12.341.28gm% and 12.341.42gm%, respectively
and at the end of experiment the Hb content reached
to
13.561.18gm%,
13.491.29gm%
and
13.401.25gm%, respectivelyCUD+A had Hb
content at zero day 12.411.48gm% which was
increased to 15.381.31gm%, at the end of
experiment. Group CUD+B had 12.291.42gm% Hb
content on zero day which was increased to
14.901.17gm% at the end of experiment. In
CUD+G and CUD+H, the Hb content at zero day was
12.271.41gm%,
12.301.39gm%
and
was
14.861.14gm% and 14.781.11gm% at the end of
the experiment, respectively.

Total Leucocyte count (TLC) in experimental mice

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Absolute lymphocyte count (AlC) of experiment of mice.

Haemoglobin content (Hb) in experimental mice

Absolute neutrophil count (AlC) of experiment of mice.

Total erythrocyte count (TEC) in experimental mice

Discussion
In-vivo study with T. baccata leaves and bark extracts
alone and in combination with CUD were carried out
in experimental mice for a period of six months. With
an observation at 15 days interval in mice, an attempt
was made to produce cancer using DEN and various
clinicohematological parameters were observed. The
body weight of mice was decreased substantially in
DEN treated mice indicating in the development of
cancer, due to DEN.Ramji and You, (1992) reported
that aflatoxin has been directly related to under
weight status in children in Benin and Togo. Bedi et
al.,(1996) reported decreased in body weight in
Guinea fowl fed on aflatoxin B1. In present study
body weight in DEN treated mice was decreased at
the end of experiment. However, there was increase
in body weight in other test groups. This study
showed that the weight loss in DEN treated group
may be due to the carcinogenic effect of DEN;
however, herbal formulations of extracts and CUD
were found to be a preventive agent against the
carcinogenic effects of DEN. DEN is already known
chemical carcinogen.Increased immunocompetence
of an individual is a very essential parameter to
prevent the development of cancers by several
mechanisms, of which the upregulation of
lymphocyte proliferation and stimulation activity,
increased macrophage activity, higher antibody
production and increased synthesis and secretion of
cytokines (IL-1, Il-2) plays significant role by
enhancing the recognition of tumor cells by the
immune cells of the body and cytotoxic activities of
the tumor killing cells, the lymphocytes. Using herbs
for cancer treatment can help the body to support its

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International Journal of Advanced Research (2013), Volume 1, Issue 5, 71-78

healing power. In the present investigation, both

doses of QC (5 and 25 mg/kg) led to a significant
decrease in the number as well as the mean area of
GST-P positive foci, TUNEL positive apoptotic cells,
p53 positive hepatocytes, and restoration of cellular
morphology. These results clearly indicate that
quercetin inhibits diethylnitrosamine-induced hepatic
preneoplastic lesions in medium-term rat liver
bioassay. In the mice given T. baccataalone and
along with CUD. The body weight either remain
constant or enhanced substantially.
These
preparations as shown in the in-vitro study were
having anti-carcinogenic effect, which might be
altering the clinoco effects of the cancer caused by
DEN.

Jemal A., Bray F., Center M.M., Ferlay J., Ward E.

and Forman D. Global cancer statistics. CA: a cancer
journal for clinicians, 2011, 61 (2): 6990.

Various hematological parameters indicated the

leukocytosis, erythrocytosis higher hemoglobin
content in treated mice with T. baccata products
along with indigenous cow urine. While, in DEN
treated mice there was leucopenia, erythropenia and
decreased heamoglobin content. These findings are
further supported by the fact that CUD had the
immunomodulatory
property
which
caused
leukocytosis leading to the control of the highly
proliferating cells through their destruction by the
white blood cells. Erythrocytosis and increased
hemoglobin content are the indication of good health
and recovery and neutralization of the effect of DEN
by T. baccata and CUD. Joshi et al., 2013
investigated that the immunomodulatory effect of
distilled
Gir
cow
urine
in
rabbits
throughhaematological parameters. The study
revealed that the values of total leucocyte count
(TLC), absolute lymphocyte count (ALC) and
absolute neutrophil count (ANC) were significantly
increased in Group II, in which the rabbits were
given Gir cow urine distillate alone and Gir cow
urine distillate with citric acid, respectively. In the
present study the ALC and ANC increased 40.31%
and 40.13% inn extracts and/or CUD treated mice in
comparison to control or DEN treated mice. Increase
in TEC and Hb content is an indication of enhanced
vitality
of
mice.
Similarly
leukocytosis,
lymphocytosis and neutrophilia are the immune cell
showing immunopoturtration, which is considered
protective against cancer and an indication of a good
prognosis (chauhan, 2005). It further needs a detailed
study for further confirmation.

Udupa A.L.,Kulkarni D.R. and Udupa S.L. Effect of

Tridaxprocumbens extract on wound healing, Int. J.
Pharmacognosy, 1995, 1: 37-40.

References

Joshi A.,Bankoti K.,Bisht T. and Chauhan R.S.

Immunomodulatory effect of Gir cow urine distillate
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Pathogenic effect of free radicals and their prevention
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Ramji C and You W. Differential sensitivity of

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***********

78