Observation of T-lymphocytes by anae staining in the clinical application of qigong

Author: Xu Hefen 1//Wang Guomin 1//Xue Huingihng 1//Zhanbg Chengming 1//Wang Junmei 2//Qi Yueqin 2
Jiangsu Provincial Institute of Traditional Chinese Medicine, Jaingsu Province, China [1] //Jiangsu Provincial Hospital of Traditional Chinese Medicine, Jaingsu Province, China [2]
Conference/Journal: 1st World Conf Acad Exch Med Qigong
Date published: 1988
Other: Pages: 52 , Word Count: 774

Malignant tumor is known as one of the common diseases, and by statistics, the patients who died of such disease are as many as 800 thousand every year, averaging one patient per 4 min. Therefore we should pay close attention to the prevention and treatment of it. In general, we have treated such patients with comprehensive treatment in order to prolong their survival periods. In recent years qigong has been used as an important integral part in the comprehensive treatment for malignant tumors, and its mechanism has been explored and discussed by observing the immunological function—the determination of T-lymphocytes by acid alpha-naphthyl(a-naphthyl) acetate esterase staining (ANAE for short), of which the following is a report.

1. Subjects observed:
272 persons were observed in total, including 72 healthy persons who practised the qigong exercise (the 1st group), 50 healthy persons (the 2nd group), 50 persons specializing in keeping bees (the 3rd group), 50 patients suffering from malignant tumors and practising the qigong exercise (the 4th group) and 50 patients with malignant tumors who did not practise any qigong exercise (the 5th group). All the malignant tumors had been identified and diagnosed by pathological biopsy.

2. Methods:
Blood smears were made of blood from their fingers or ear lobes. After being dried, they were put into a kind of solution for 20-30 sec. for fixation. Then they were taken out, rinsed with distilled water and dried. They were put into a kind of incubative solution for 2 hrs. Then the precipitate was rinsed with the fountain water, dried in frozen condition and re stained with methyl green for 1-1.5 min. Lastly, the smears were observed under the oil immersion microscope and 200 Iymphocytes should be counted. Of them those having dark red minute particles were T-lymphocytes, while those not having such particles were

Fixing solution:
Na2HPO4 20 mg
KH2PO4 100 mg
H2O 30 ml
Acetone 45 ml
40% Formaline 25 ml PH 6.6

Incubative solution:
(1) Parafuchsin solution: 2g of parafuchsin mixed with 50 ml of 2N HCl, reserved in a refrigerator of 4°C.
(2)4% NaNO3 solution: 0.4g of NaNO3 dissolved in 10 ml of distilled water (to be made up when it is to be used). [Editor's note: we assume NaNO[sub 3] to be correct although the orginal text indicates NaNO:]
(3) Drip 3 ml of 4% NaNO3 solution slowly into 3 ml of parafuchsin solution, let them mixed fully and set the mixture aside for one minute, to be made up at the last moment).
(4) 2% alpha-naphthyl-acetate solution 2g of alpha-naphthyl acetate dissolved in 100 ml of methyl glycol, stored in a refrigerator of 4°C and kept in a dark place.
(5) M/l5 PH 7.6 phosphoric acid buffer solution:
Solution A: 100 ml containing 9.08g of KH2PO4 and water.
Solution B: 100 ml containing 9.47g of Na[sub 2]HPO[sub 4] and water.
Mix 13 ml of solution A with 87 ml of solution B when they are to be used.
(6)Make up the incubative solution when it is to be used.
Mix fully 89 ml of M/15 PH 7.6 phosphoric acid buffer solution and 6 ml of parafuchsin NaNO3 solution. Drip slowly 2.5 ml of 2% alpha-naphthyl-acetate solution into the above solution to make a perfect mixture with PH 5.6-6.4.
Methyl green staining solution: 0.5g of methyl green dissolved in 50 ml of distilled water, assisted by heating.

3. Result and Discussion:
The value of X±SD of ANAE determination of the 1st group was 74.9±11.6l%, while that of the 2nd group was 65.60±8.9%. By comparison, the difference was significant (P< 0.01). The value X±SD of the 4th group was 69.2±12.77%, while that of the 5th group was 42.82±7.09%. By comparison, the difference was significant (P<0.01) And the value of ANAE of the 3rd group was 76.87±11.06%. From this observation, we could find that the mean value of the 3rd group was obviously higher than that of the 2nd group or the 5th group.

The results mentioned above showed that the value of ANAE of these persons who had practised the qigong exercise or who specialized in keeping bees was obviously higher than that of the healthy or those patients who did not practise the qigong exercise. Besides, the value of ANAE of the cancer patients who practised the qigong exercise was much higher than that of those cancer patients who did not practised the qigong exercise.

The results of observation suggest that the qigong exercise may enhance and promote human immunological function, resulting in strengthening the defensive resistance and recovering from such diseases as malignant tumors. Therefore it is suggested that we should continue to carry out systematic and intensive research on the relationship between the qigong exercise and the immunological function of the human body, with which we can further elucidate and make the mechanism of treatment of malignant tumors by means of the qigong exercise understood.