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| Position Now: English version - Experiment
researches |
Experimental
researches on animals- material development
and method of the experiment |
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- Development of animal models:
(1) Model of femoral head necrosis: rats
whose ages were all less than 10 days old. Their forelimbs were
cut off from above the elbow joints. Their tails were cut off
from the bass. After these cuts, the rats continued to be fed by
mother rats. After one month, the average body weight of these
rats was more than 150g. From then, on the rats were given
Prednisone: 20mg/kg, qod, 3 months. (2) Model of osteoporosis:
female rats whose weights were all 200±20g had remove both
ovaries to simulate rats that were in the condition of extremely
low estrogen after menopause.
- The medicine observed or studied:
Chengzai pill
developed and supplied by Professor Huang (processed by Jilin
pharmaceutical factory). The authorization number was Jilin-Health-Drug-Self
made- 92002.
- Experimental groups, dose, and means of medication:
Normal contrast group, negative contrast group, and
“Chengzai pill” therapeutic group were established for studying
the effect of the medicine used for femoral head necrosis.
Besides the three groups above, another group, the alphaD3
group, was established for studying the effect of the medicine
used for osteoporosis.
(1) Normal contrast group: Normal contrast group of osteoporosis
study were adult female rats. Normal contrast group of femoral
head necrosis study were the little rats like the models of
femoral head necrosis. However, these rats were not managed.
(2) Negative contrast group: two kinds of models with no other
medications .
(3) “Chengzai pill” therapeutic group: the models that
were given “Chengzai pill”. For femoral head necrosis study the
dose was 8 pills qd 3 months. For the osteoporosis study the
dose was 8 pills qd 3 months. For the once using Prednisone,
“Chengzai pills” were given as in the femoral head necrosis
study. After both ovaries were removed, “Chengzai pills” were
given 3 days later for the osteoporosis study.
(4) Vit-D contrast group: Alpha D3 was used qd for
3months. The dose was 0.0042 equal to the adult aequum.
- Indexes of observation and detecting method
(1) Osseous density all over the body: Measured by
apparatus of BMD（Lunar DPX）under the narcosis of Phenobarbital.
(2) Osseous density of femur: One side of femur was taken out
after being put to death by bloodletting through abdominal aorta
under the narcosis of Phenobarbital. Density of the femur was
detected by apparatus of BMD.
(3) Detection of intensity and rigidity: Intensity and rigidity
were detected by electronic versatile test – machine (type:
WD-1, produced by Changchun metalloid test-machine fatory) after
the detection of density.
(4) Moist weight, dry weight and ash weight of femur: Moist
femur was weighed by electronic balance after detection of
intensity and rigidity. Then it was dried under the temperature
of 60 ℃ for 4 hours, and weighed. Finally it was incinerated in
Ma’s asher, and weighed.
(5) Capillary distribution of femoral head: Inject 5ml ink
through femoral artery
under the narcosis of Phenobarbital. The femur on the injection
side was taken out and immersed in Formaldehyde for fixation.
Then it was dehydrated by alchohol and embedded in Methyl
methacrylate. Finally it was cut into 0.3mm-0.5mm bone slides
which could be observed under the light microscope. The form and
distribution of capillaries on the slides could be confirmed.
The proportion of capillaries in an unit area also could be
decided.
(6) Fat deposition in the osseous tissue of femoral head: The
other femoral head was taken out and fixed in 70%-90% alcohol.
Then it was frozen, sectioned and stained with Lillie’s oil red
O. Finally the distribution of fat was observed under light
microscope. The proportion of chondrocytes and osteocytes which
were stained was calculated.
(7) Observation by scanning electron microscope: Superficial
form and fine structure of femoral head were observed under
scanning electron microscope.
(8) Level of estrogen determination: The serum was taken out and
the level of estrogen was determined by radioimmunoassay. ( the
reagents were produced by Northern Immunoreagent Institute of
Beijing)
(9) Level of 25(OH)D3 determination: The level of 25(OH)D3 was
determined by competitive protein combined assay.
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