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Observation of Antiaging
Effects of Vital Cell in Rabbits

Materials: 20 4-month-old, healthy purebred New Zealand rabbits, 10 males and 10 females each weighing 2kg. (Supplied by Shanghai Laboratory Animal Center of China Academy)

Grouping: Rabbits were divided randomly into two groups of 10, labeled Group A (the administration group) and Group B (the control group).

Feeding: Every rabbit in both groups is fed in a separated cage in the same room. All are observed under room temperature. The feeding of rabbits is strictly defined according to time and quantity.

Observation Index and Methods

Method: After 2 years of treatment researchers randomly selected one male and one female rabbit from each group (A -treatment and B-control). Researchers also selected one male and female rabbit from a group of 6-month-old, healthy purebred New Zealand rabbits weighing 2kg each. This group was labeled Group C (young, healthy controls).

Researchers randomly numbered all 6 rabbits and, after etherizing them, obtained tissue samples from for examination with both optical and electron microscopes. Tissues were prepared for optical microscope by fixing with formalin, paraffin section, HE staining, weight elastic fiber staining on the myocardium and coronary arteries, and NAP staining fixed by acetone for womb tissues.
Tissue samples for the electron microscope were fixed with 2.5% sporicidin, embedded with expoxide resin, prepped into ultra-thin sections and stained with acetate uranium and citrialuminum.

Observation Index: Researchers examined tissue samples from the thymus glands, hearts, lungs, livers, spleens, kidneys, brains, wombs and testes/womb of the 6 rabbits from Group A, B, and C under optical microscope.

Researchers examined tissue samples from the livers of all 6 rabbits and testes of the 3 male rabbits from Group A, B, and C under electron microscope.

Results of Observation with Optical Microscope

Thymus gland: The weight of the thymus gland of the rabbits in Group A was 2g. The atrophy of the thymus gland was relatively slight. The weight of the thymus gland of the rabbits in Group B was 1g, the cells of the thymus gland decreased and adipose tissues overgrew. The weight of the thymus gland of the rabbits if Group C was 3g and the tissues and structure remained normal.

Thymus - Old rabbit
Thymus with severe atrophy,
heavy fatty infiltration
Thymus - Treated rabbit
Firm mass, only slight
signs of atrophy

Heart: There were very few micro-fiber foci in the myocardium of the rabbits in Group A. The pathological changes in the coronary arteries were not obvious, and the elastic intima was clear. The myocardium of the rabbits in Group B was fiberized. There were deposits of lipofuscin. The collagen fibers of the coronary arteries overgrew, the intima became thickened irregularly and the elastic intima split and proliferated. The myocardium and coronary arteries of the rabbits in Group C kept normal.

Heart - Old rabbit
Sclerosis of coronary artery,
thickened intima, fibrosis
Heart - Treated rabbit
Slight myocardial fibrosis and
unclear coronary sclerosis

Lung: There were no obvious pathological changes in the blood vessels of the lungs of the rabbits in Group A. The wall of the arteriole of the lungs of the rabbits in Group B thickened and the arteriole became narrower. The blood vessels of the lungs of the rabbits in Group C were normal.

Lung - Old rabbit
Thickening of small arteries,
narrowing of arterioles
Lung - Treated rabbit
No thickening or damage
to small artery walls

Liver: There was a little blood stagnation of the liver in the rabbits of Group A, but the atrophy of the hepatic cells was not very obvious. Very little lipofuscin was found. There was obvious blood stagnation and atrophy of the liver and obvious atrophy of the hepatic cells in the rabbits of Group B. Deposits of lipofuscin could be found. The livers of the rabbits in Group C were normal.

Liver - Old rabbit
Stagnant blood, lipofuscin
deposits in cells
Liver - Treated rabbit
Absence of blood stagnation,
no lipofuscin in cells

Spleen: There was no blood stagnation of the spleen in Group A. The walls of the central arteries of the splenic corpuscles thickened slightly. There was blood stagnation of the spleen in Group B, and the walls of the central arteries of the splenic corpuscles thickened thus causing the arteries to become narrower. The rabbits in Group C were normal.

Spleen - Old rabbit
Vessel wall thickening of central
arteries in splenic corpuscle
Spleen - Treated rabbit
No vessel wall thickening of
arteries in splenic corpuscle

Kidney: There was no obvious unit decrease of the kidney of the rabbits in Group A. Under the view of 10 lower diameters, 300 glomeruli could be found. However, there was unit decrease of the kidney of the rabbits in Group B. Under the view of 10 lower diameters, some 230 glomeruli could be found. The walls of arteriole in the kidney thickened and became narrower. In the rabbits of Group C, about 330 glomeruli were found under the view of 10 lower diameters.

Kidney - Old rabbit
Decrease in nephrons
with few glomeruli
Kidney - Treated rabbit
Slight decrease in nephrons

Brain: The soft meninx of the rabbits in Group A did not thicken. The blood vessels expanded, but there was no obvious blood stagnation. The soft meninx of the rabbits in Group B, thickened, and the blood vessels of the lower part of the arachnid membrane expanded. There was evidence of blood stagnation. Part of the blood vessels of the brain became spuriously calcified. The rabbits in Group C were normal.

Brain - Old rabbit
Calcification of arterial vessels,
thickening of soft meninx
Brain - Treated rabbit
No thickening of soft meninx,
slight expansion of vessels

Uterus: There was the enlargement of uterus in the rabbits of Group A, and the intima thickened. AKP staining showed (-) for the rabbits of Group A. There was atrophy of the womb in the rabbits of Group B. The womb became narrower, and the intima was thinner. AKP staining showed (+).

Uterus - Old rabbit
Thinning oftunica intima
of the uterine lining
Uterus - Treated rabbit
No thinning of tunica intima

Testis: There was no obvious atrophy of the spermaduct in the rabbits of Group A, and there was a large amount of sperm. There was atrophy of the spermaducts in the rabbits of Group B and the sperm cells of various kinds were degenerated, reducing the number of sperm.

Testis - Old rabbit
Atrophy of the seminiferous
tubules, reduced sperm
Testis - Treated rabbit
No atrophy of the seminiferous
tubules, healthy sperm

Results of Observations with Electron Microscope

Liver: In Group A, the nuclei were either circular or oval, while the nuclear membrane was clear. The nucleoli had a thread-like structure. The nuclear chromatin was well distributed. The shape and volume of the mitochondria in Group A were similar to those in the healthy young rabbits. They were thick with numerous endoplasmic reticulum. Reticulation enlargement was not apparent.

In Group B, the nuclei had an irregular shape. The nucleoli were separated and had lost their thread-like structure. Around the nucleoli, the chromatin had increased. Cytoplasmic mitochondria were swollen and shortened, and their shape was irregular. The dilatation of endoplasmic reticulum decreased. Lipofuscin deposits could be observed.

In Group C, the mitochondria were slightly enlarged. Their other structures were normal.

Testis: In Group A, no atrophy of convoluted tubule was observed, and there were no apparent degenerative changes in spermatoblasts of different levels. The nuclei or cell organs had a clear structure.

In Group B, the convoluted tubule was found to have shrunk. Spermatoblasts of different levels and sterol cells showed degenerative changes. Nuclear chromatin condensed. Denaturation of intracytoplasmic cell organs and their obscure structure could be observed.

Results

In Group B, thymus glands, womb and testis displayed noticeable shrinkage. Coronary sclerosis, myocardial focal fibrosis, lipofuscin deposits and nephronal decrease were common. The use of an electron microscope revealed patent degenerative changes in the nucleoli and mitochondria of hepatic cells and convoluted tubule cells, accompanied by lipofuscin deposits.

In Group A, fewer pathologic changes were apparent. In comparison to Group C, the organic functions in Group A were normal.

* These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.

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