Chinese Medicine can enhance immune system

With the continuous advancement of scientific research means, the understandings for our body are advancing continuously. New findings have made clear that the immune function is not only associated with the infectious diseases but also associated with non-infectious diseases closely. Such as, cancer, an uncontrolled cellular proliferation diseases associated with the immunity. Therefore, seeking the approaches to enhance our immunity have been becoming the research focus and have gotten great many fruits.

Based on the unique philosophy and independent medical theory, with an abundant natural medicinal resources, Chinese drugs have been demonstrating amazing therapeutic effects and attracting more and more attention worldwide. And great many Chinese drugs have been revealed therapeutic mechanisms in the level of biochemical and pharmacological. They are ready to be used clinically.

Modern researches showed that Chinese drugs contain variety of immunoreactive substances, such as polysaccharides, saponins, alkaloids, volatile substances and organic acids, etc. They are a kind of immune enhancer. Their immunoregulatory role is mainly through activation of reticuloendothelial system, so that significantly increase the IL-content, and the level of antibodies and complement, activate the macrophages and T, B lymphocytes, and enhance cellular immunity and humoral immunity, thereby enhancing the body immune system and the capacity of anti-diseases.

More amazingly, Chinese drugs can intellectually take the bidirectional role for those under different immune status. That is, for those in the lower immune function play a catalytic role, for those in the over immune function play a disincentive role, and for those in healthy condition stop any action consciously.

The article is trying to demonstrate the effects of Chinese drugs on enhancing the immunity by some research articles. It is only a rough glance.

2. The effects of Chinese drugs on the development of immune organs

2.1. Effect of Traditional Compound Chinese Medicine on the Ultrastructure of Immune Organs in Mice Bearing S180 Cancer

Methods: 55 mice divide into 5 groups, includes positive and negative control groups. The 2 10^7 S180 cells were resuspended in 0.2 mL PBS solution and were injected subcutaneously into the right haunch of mice. The mice bearing S180 sarcoma in three groups were administrated with compound Chinese drugs with doses of high (0.5g/mL), middle (0.25g/mL) and low (0.125g/mL) by gavages. The index of spleen and thymus were calculated. The proliferation of the spleen and thymus lymphocyte was observed by electron microscope.

Results: Compound Chinese drugs can increase the index of spleen and thymus and the capacity of proliferation of spleen and thymus lymphocyte. The proliferation is significant in high and middle dose group. The appetite of mice was improved and the weight was gained.

Conclusion: Compound Chinese drugs have significantly inhibitory effect on S180 mouse sarcoma by improving immunologic function.

2.2. The effect of Chinese drugs on immune system and weight gain of Rabbits

Methods: Normal reared rabbits were divided into treatment group and the control group. Chinese drugs accounted for 2% of rabbit diet in the treatment group.

Results: the average daily weight gain of rabbits in the test group is 7.13% higher than that of control group (P<0.05). Compared with the control group, in treatment group, the index of immune organs of spleen, thymus, mesenteric lymph nodes, sacculus rotundus and appendix were separately increased 12.93% (P

Studies have shown that adding Chinese drugs in the diet of baby rabbit can significantly improve their immune system, and improve the feed utilization by lowering the feed/gain.

Conclusion: Chinese drugs can enhance the immune system and weight gain for rabbits.

2.3. Effect of Taraxacum Polysaccharides on Immune Organs in Mice

Methods: 30 mice divided into Taraxacum group and control group, 15 mice each group. Dandelion polysaccharide solution gavaged for mice in Taraxacum group and equal weight saline solution gavaged for mice in control group, all administrated for 7 days.

Results: Visual observation

The color, shape of the spleen and thymus is normal in each group, no visible pathological change. Compared with the control group, in treatment group, the spleen, thymus are full of shiny, bulky, obvious changes can be seen.

The spleen index and thymus index

Groups	Spleen index	Thymus index
Control group	3.13±0.23	4.27±0.37
Treatment group	3.95±0.31*	5.54±0.48*

Note： “*” means the difference is significant (P<0.05).

The organizational structure of the spleen and thymus

The slices of mouse immune organs were observed by optical microscope, compared with the control group, for the mice in treatment group, the boundaries of the spleen white pulp and red pulp is significantly clear. The area of white pulp was increased. The lmphoid nodule and splenic corpuscle was increased. No abnormal changes on the organizational structure were found on the slice of mice immune organs in each group.

Conclusion: Dandelion polysaccharide can promote growth and development of the spleen and thymus of mice to enhance immune function.

2.4. The effects of Chinese drugs compound on immune organs of chickens

Methods: 60 7-day-old AA Chickens were randomly divided into Chinese drug group and control groups of 30 each. 0.5% Chinese drugs compound added into the Chicken diet for drugs group. The control group gives normal diet. Two groups were administrated for 14-day.

Results: The weights and index of thymus, bursa of Fabricius and Spleen are as following.

Organs	Groups	Weight/g	Index/×10-3
Thymus	Drugs	5.12±0.027a	3.19±0.217a
	Control	3.38±0.521b	2.67±0.091b
Bursa of Fabricius	Drugs	4.08±0.029a	3.92±0.005a
	Control	2.51±0.142b	2.44±0.012b
Spleen	Drugs	1.98±0.011a	1.89±0.311a
	Control	1.23±0.073b	1.34±0.771b

From above table, in drugs group, the weight and index of thymus, bursa of Fabricius and spleen were significantly higher than those of control group (P<0.01).

Conclusion: Chinese drugs can significantly increase the weight and index of thymus, bursa of Fabricius and spleen of Chicken.

2.5. Effect of Chinese drugs on immunodeficiency of insomnia mice

Methods: Set a control group first, and the rest of mice were established mice insomnia model by chronic restraint stress for 7 days, then divided into 2 groups, treatment group and model group. The treatment group intraperitoneal injection of Chinese drugs 40 ml/kg per day, and the model group were given same dose of normal saline.

Results: Compared with control group, in the model group, the body weight of mice decreased, pentobarbital sleep time was significantly shortened, immune organ weight, the percentage of spleen CD3+、CD3+CD4+CD8-、CD3+CD4+CD8-/CD3+CD4-CD8+(CD4+/CD8+)、CD3+CD25+ and the capacity of ConA-induced splenic lymphocyte transformation were all significantly decreased (P＜0.05). In treatment group, the degree of mice weight loss reduced significantly, immune organ weight, the percentage of spleen CD3+、CD3+CD4+CD8-、CD3+CD4+CD8-/CD3+CD4-CD8+(CD4+/CD8+)、CD3+CD25+ and the capacity of ConA-induced splenic lymphocyte transformation were all significantly increased (P＜0.05). And are close to normal levels (P>0.05).

Conclusions: Insomnia may lead to dysfunction of cellular immunity in mice. Chinese drugs is effective to restore the cellular immune function of mice in insomnia model.

3. The effects of Chinese drugs on the specific immune

3.1. The effects of Chinese drugs on cell-mediated immunity

3.1.1. Effects of Chinese drugs’ immunomodulator on cellular immune function in mice

Methods: 72 mice divided into 6 groups randomly. Group 1 was control group in normal diet. Group 2 was immunosuppressant group, injection of cyclophosphamide 50 mg/kg/day, for 7 days. Group 3 and 4 was the Chinese drugs’ immunomodulator groups administrated by drinking once daily for 15 days. Group 5 and 6 was the general Chinese drugs groups administrated as same as the group 3 and 4.

After 4 days Chinese drugs administration, group 4 and 6 was injected cyclophosphamide, the dose and the course was as same as group 2.

Results: Mice in immunosuppressant group 2, the A570 value of the splenic lymphocyte transformation was significantly low than those in the control group (P

In the test of mice splenic lymphocyte transformation, whether adding concentration of 50 mL/L or 100 mL/L of serum of each group, The A570 value of mice in group 4 and 6 was significantly higher than that of any group (P<0.05). The A570 value of mice in immunosuppressant groupm was significantly low than those in the control group (P

Conclusion: Chinese drugs have significant effect of immune regulatory on the mice in abnormal immune function. There is no effect on normal mice.

3.1.2.The effects of Chinese drugs compound on celular immune function of chickens

Methods: 60 Chickens, 7-day-old, were randomly divided into Chinese drugs group and control groups of 30 each. 0.5% Chinese drugs compound added into the Chicken diet for drugs group. The control group gives normal diet. Two groups were administrated for 14-day.

Results: The formation rate of T lymphocyte E rosette ring

The test results are as following table. The mice in Chinese drug group, the formation rate of T lymphocyte E rosette ring was significantly higher than those of control group (P<0.05).

Test results of the formation rate of T lymphocyte E rosette ring

Groups	Brfore drugs (X±SD)	After drugs (X±SD)
Drugs	34.12±3.56	47.63±6.01a
Control	33.67±2.93	33.88±4.65b

The formation rate of B lymphocyte EAC rosette ring

The test results are as following table. In Chinese drugs group, the formation rate of B lymphocyte EAC rosette ring was significant higher than that of control group (P<0.01).

Test results of the formation rate of B lymphocyte EAC rosette ring

Groups	Brfore drugs (X±SD)	After drugs (X±SD)
Chnese drugs	41.14±5.91	50.04±6.43a
Control	42.36±6.33	40.19±5.81b

The formation rate of erythrocyte rosette

The test results are as following table. In Chinese drugs group, the formation rate of E-C3bR rosette was significant higher than that of control group (P<0.01).

The test results of the formation rate of E-C3bR rosette

Groups	E-C3bR rosette rate
Chinese drugs	19.50±3.40a
Control	13.50±1.86b

Conclusions: Chinese drugs compound can increase the formation rate of T lymphocyte E rosette ring, the formation rate of B lymphocyte EAC rosette ring and the formation rate of erythrocyte rosette. The results showed that the Chinese drugs compound can enhance the level of the cellular and humoral immunity.

3.2. The effect of Chinese drugs on the humoral immunity

3.2.1.The effects of Chinese drugs on the level of Newcastle disease antibody

Methods: 60 Chickens, 7-day-old, were randomly divided into Chinese drug group and control groups of 30 each. 0.5% Chinese drugs compound added into the Chicken diet for drugs group. The control group gives normal diet. Two groups were administrated for 14-day.

Results: In Chinese drugs group, the level of Newcastle disease antibody was significant higher than that of control group (P<0.01).

Groups	level of antibody
Chinese drugs	5.75±0.20a
Control	4.42±0.26b

Conclusion: Chinese drugs can significantly increase the level of Newcastle disease antibody of Chicken.

3.2.2. The influence of astragalus on the expression of interleukin-2 and soluble interleukin-2 receptor in experimental obstructive jaundice in rats

Objective: To investigate the protective effect of astragalus on the injury of the immune system in obstructive jaundice by establishing obstructive jaundice rat model and by intervention in administering astragalus agents, thus offering a potential therapeutic strategy to obstructive jaundice diseases.

Methods: 30 male Wistar rats were divided into three groups randomly and on the average. A: sham operation group; B: obstructive jaundice (OJ) group; C: OJ+Astragalus group. Ten days later the rats were sacrificed and the serum was obtained to detect the ALT, bilirubin, IL-2 and sIL-2R in each groups.

Results: Compared with groups A and C, the IL-2 level in group B decreased and sIL-2R level increased, with marked evidence of injury to the immune functions. There was statistical significant difference between group B and C (P<0.05 and P<0.01, respectively). The hepatic functional parameters in group C were lower than those in group B (P<0.05).

Conclusion: Astragalus can enhance the immune function by increasing the level of IL-2 and decrease the level of sIL-2R.

3.2.3. The effects of Chinese drugs on Acquired immune deficiency syndrome (AIDS)

In Tanzania, there were eight cases of serum antibody turning to negative among the AIDS patients that have been treated by Chinese specialists with Chinese drugs. Moreover, they are unchanged after 49 months’ continuous tracking.

The eight cases of AIDS are half men and women. There is a young man AIDS patient, 20, named J.L. For two months he have been fatigue, shortness of breath, skin itching, throat discomfort and the Karnofsky score was 80 points, ELISA test is positive. After treatment by Chinese drugs, fatigue symptoms quickly eliminated. During the treatment, all the other symptoms have disappeared. Review after 87 days, found his ELISA was negative.

The facts showed that the process of AIDS pathogenesis is reversible through the proper treatment and Chinese drugs have therapeutic effect on AIDS. It provided a new hope for the treatment of AIDS.

4. The effects of Chinese drugs on non-specific immunity

4.1. Leukocyte and monocyte-macrophage system

4.1.1. Astragalus injection treats Leukopenia

Case selection: 67 hospitalized patients, the interleukin value are all<4.0×109/L with the symptoms of varying degrees of dizziness, fatigue, weakness, susceptible to colds and so on. They were randomly divided into two groups.

Treatment group: 35 cases, 7 male, 28 female, aged 18 to 50 years old, average 33-year-old. Leukopenia reasons: radiotherapy and chemotherapy 4 cases, immune factors 4 cases, viral infection 14 cases and unexplained 13 cases.

Control group: 32 cases, 9 male, 23 female, aged 20 to 56 years old, average 37-year-old. Leukopenia: radiotherapy and chemotherapy 6 cases, immune factors 4 cases, viral infection 7 cases, 15 cases of unknown causes.

Treatments: Treatment group: Astragalus injection 40 ml ( Pharmacognostic containing 10 g/ml), adding 250 ml of intravenous glucose or saline, 1 times/d, 15 d for a course of treatment, observation for two courses of treatment. Control group: Oral batyl alcohol, leucogen, intravenous energy mixture, 15 d for 1 course of treatment, observation for two courses of treatment.

The standard of efficacy: Effective markedly: WBC returned to normal or increase 1 times than that of pre-treatment. Effective: WBC values increases ＞0.5 × 109/L than that of before treatment. Invalid: WBC values increase ＜0.5 × 109/L than that of before treatment.

Results: ① Treatment group: effective markedly 7 cases, 20.0%, effective 24 cases, 68.5%, ineffective: 4 cases, 11.0%. ②Control group: effective markedly 4 cases, 12.5%, effective 17 cases, 53.0%, ineffective 11 cases, 34.4%. The total effective rate in treatment group was 88.5%, in control group was 66%. Comparing the two groups were statistical significant difference (P<0.05).

Changes in white blood cells: Before treatment the WBC values were lower than normal in both groups. Comparison between the two groups was no significant differences (P＞0.05). After treatment, the treatment group was from 2.12 ± 1.22 × 109/L increased to 3.66 ± 1.02 × 109/L (P＜0.05). The control group was from 2.14 ± 1.08 × 109/L increased to 2.98 ± 1.20 × 109/L, the difference between the two groups was significant.

Clinical symptoms: The treatment group, by the end of a course of treatment, dizziness, weakness, etc. has been noticeable improved. However, for the control group, often needs more than two courses of treatment, these symptoms were only partial remission.

Conclusions: The effective rate reached 88.5% in treating Leukopenia by Astragalus injection and it was 66% by Western drugs. Astragalus injection was made from the pure natural Chinese drugs with lesser negative effect. It is a promising alternative drugs for treating Leukopenia

4.1.2. The effects of anti-virus preparation of Chinese drugs on immune cells

Methods: Mice (weight 25~28g, half male and female) were randomly divided into two groups, Test groups of mice gavage 0.4 ml anti-virus mixture daily. The mice of control group fed with 0.4 ml normal saline daily.

The staining rate of T lymphocytes esterase and the phagocytic rate of celiac macrophage were measured before administration (0 d) and after administration of 7 d, 14 d.

Results: The staining rate of T lymphocytes esterase

The test results were as following table. After administration 7 d, the number of mature T lymphocytes increaesed rapidly in mouse blood of test group. It was significantly higher than the control group. After 14 d the number of mature T lymphocytes of mice blood was no longer continues to increase in both groups. However, in the mice blood of test group, the number of mature T lymphocytes remained in a high level.

The staining rate of T lymphocytes esterase of mouse blood before and after administration

Groups	Time/d	Mice number	Esterase staining %
Test	0	5	37141±3194a
Control	0	5	37151±1181b
Test	7	10	48198±3152c
Control	7	10	41106±1177d
Test	14	10	48181±2116e
Control	14	10	42108±2120f

The phagocytic rate of celiac macrophage

The test results were as following table. After administration 7 d, the phagocytic activity of celiac macrophages of mice in test groups have been significantly higher than those of control group. In 7 d of control group, the phagocytic activity of celiac macrophage of mice was increased, but still lower than that of test group. After 14 d, phagocytic rate of mice in control group was no longer continued to grow. However, in test group, phagocytic rate of celiac macrophages of mice reached 65,164%, that is significantly higher than 37,166% of control group (P<0.01).

The phagocytic rate of mice celiac macrophage before and after drugs administration

Groups	Time/d	Mice umber	Phagocytic %
Test	0	5	28119±4101
Control	0	5	27168±4164
Test	7	10	55147±4113
Control	7	10	39163±4100
Test	14	10	65164±7160
Control	14	10	37166±4119

Conclusion: The anti-virus preparation of Chinese drugs can significantly increase the the number of mature T lymphocyte and the phagocytic rate of mice celiac macrophage in mouse blood.

4.1.3. The effects of compound Chinese drugs on promoting T lymphocyte proliferation

Methods: Separate T-lymphocyte from blood of SD rat, cultured in vitro. Install normal cell control group, positive-drug control group, transfer factor group, compound Chinese drugs high dose group, middle dose group and low dose group.

Added the drugs to the culture medium after 1 day, then continue to culture 2 days and take Am-Blue into 96 orifice plates, and then cultured 4 hours in dark. Use enzyme-labelling measuring instrument to measure and analyse proliferation of T-lymphocyte after 3 days.

Results: The test results are as following table. The OD value in 3 dose Chinese drugs groups was increased significantly than those of normal cell control group and positive-drug control group P＜0.01. In Chinese drugs high dose group increased significantly than those of middle dose and low dose group P＜0.01. Transfer factors group increased significantly than those of normal cell control group P＜0.01.

The test results of OD value of each group

Groups	Cases	OD value	P Value
Normal cells	10	0.04±0.22	<0.01
Positive-drugs	10	0.11±0.24	<0.01
Transfer factors	10	0.13±0.41	<0.01
Chinese drugs low	10	0.20±0.38	<0.01
Chinese drugs middle	10	0.40±0.31	<0.01
Chinese drugs high	10	0.54±0.12	<0.01

Conclusion: Compound Chinese drugs can promote the T-lymphocyte proliferation with dose-dependent.

4.2. The effects of Chinese drugs on cytokines

4.2.1.The effects of Chinese drugs on cytokines in arthritis rats

Methods: 40 wistar rats divided into 5 groups randomly. Group 1 is the blank control group, group 2 is the model control group, group 3 is the MTX group, Group 4 and group 5 is the Chinese drugs groups. For rats in group 2, 3, 4 and 5 were made of animal models of rheumatoid arthritis (RA) by injection of Freund’s complete adjuvant in the left foot, each 0.1 ml (containing BCG 10mg/ml). Group 1 was saline injection. After 1 week of proinflammatory starts medication, gavage MTX 5mg/kg weekly and Chinese drugs 18g/kg daily. All group’s rats was administrated for 3 weeks. Then, 5 ml serum was isolated from the heart blood. The TNFα measured by radioimmunoassay, the IL-1β measured by enzyme-linked immunosorbent assay method. The routine pathological examination for rats’ knee carried out after fixed by 10% formaldehyde.

Results: Model control group, both IL-1β and TNFα were higher than the blank control group, it indicates that TNFα and IL-1β plays an important role in the pathogenesis of RA. After the medication of MTX and Chinese drugs, the TNFα and IL-1β was reduced.

In model control group, the hyperplasia of inflammatory cells and edema can be seen in the knee Joint synovium of the rats. After 3 weeks medication, the inflammatory cell infiltration and the edema reduced in Joint synovium of rats.

Conclusion: Chinese drugs can significantly reduce the IL-1β and the TNFα for the mice in rheumatoid arthritis.

4.2.2. The effects of Chinese drugs on immunoregulation function of dendritic cell (DC) in peripheral blood of patients with psoriasis vulgaris

Methods: The DCs in peripheral blood of patients with psoriasis vulgaris were cultured in vitro，then they were treated by Chinese drugs decoction. The phenotypic changes of these cells were detected by flow cytometry，the proliferation state induced by lymphocytes was observed by MTT method and the changes of cytokine secreted by DC were observed by ELISA.

Results: After treated by Chinese drugs decoction, the expression of CD83 was significantly increased. However, the expression of CD86 and the stimulated ability of mixed lymphocyte proliferation were inhibited at some extent. The generation of Th1 type cytokines was decreased obviously. However, the generation of Th2 type cytokines was increased.

Conclusion: Chinese drugs have certain regulatory effect on the antigen presentation in patients with psoriasis vulgaris, and have inhibition effect on the secretion of Th1 type cells, but have promoting effect on the secretion of Th2 type cells.

4.3. The effects of Chinese drugs on natural killer cells (NKC)

Mordern researches show that Chinese drugs can promote the immunization for immunocompromised mice and can significantly enhance the Killing effect of NK cells on target cells.

4.3.1. The effects of Chinese drugs on Natural Killer Cell and Activity of Interleukin-2 of P388 Leukemia Mice after chemotherapy

Methods: Set a blank control group, then 40 mice were transplanted by strains of P388 lymphocytic leukemia tumor and were divided into leukemia model group, chemotherapy group, chemotherapy plus low-dose Chinese drugs group and chemotherapy plus high-dose Chinese drugs group.

Results: Chemotherapy can lower the NKC and IL-2 activity of P388 leukemia mice significantly than those in blank control group (P<0.01). After 2 weeks Chinese drugs treatment, the lowered activity of the NKC and the IL-2 can obviously be improved with dose-dependent.

Conclusion: Chinese drugs can be used as biological response modifier for chemotherapy-induced immune suppression of malignant cancers clinically.

4.3.2. The effects of Chinese drugs on immunoregulatory for esophageal and gastric cancer with chemotherapy

Case selection: 50 Hospitalized patients, 36 male, 14 female, aged 17-75, 25 cases of gastric cancer and 25 cases of esophageal cancer. They were randomly divided into control group of 25 cases and Chinese drugs group of 25 cases.

Treatments: For control group, hydroxycamptothecin 10 mg, fluorouracil 500 mg, cisplatin 20 mg, Calcium Folinate 100 mg, 4 drugs combined for 5 d, every 4 weeks for one cycle, two cycles for a course of treatment, each patient give one course of treatment (about 60 d). For Chinese drug group, besides above treatment, at the same time, oral Chinese drugs once daily.

Results: Compared with the control group, in Chinese drugs group, except CD8 was no different, all other indicators were better than those of control group. That CD3，CD4，CD4/CD8，AgNoRS and NK cells were significantly different (P>0.05). The activity of T lymphocytes and NK cells increased. Chinese drugs have following advantages:

The activity of T lymphocytes and NK cells increased shows that Chinese drugs can enhance the immune function.

Increase the efficacy of chemotherapy drugs. Marked effective was 40%, effective was 88%.

③ Extend survival time. 6 months survival rate was 72%, 1-year survival rates was 32%.

④ Improve the life quality of patients.

Conclusion: Chinese drugs can enhance the immune function for the patients of esophageal and gastric cancer with chemotherapy.

4.3.3. The effects of Rhubarb on Acute Toxicity and Modulation of Innate Immune in Mice

METHODS: After LD50 test, the mice were randomly divided into four groups: the 0.9％ saline control group and 3 Chinese drugs groups in dose of 1000 mg/kg, 2500 mg/kg and 5000 mg/kg separately, gavages once daily for 10 days. The complement activities and the erythrocyte CR1 adherence function were measured respectively by trace fast plate complement activity assay and complement activating yeast rosette test. NK cells killing activities were measured by improved MTT method.

RESULTS: The LD50 of rhubarb for mice was about 8043 mg/kg. Rhubarb can significantly increase the killing activity of NK cells and erythrocyte CR1 adherence function, compared with the control group there were statistical significant differences (P<0.05). The total complement activity was lower than that of control group.

CONCLUSION: Rhubarb can promote immune cell proliferation and enhance cell immunity. It can increase the NK cells killing activity and Erythrocyte CR1 adherence activity. It could be used as an adjuvant therapy.

5. The immunosuppressing effects of Chinese drugs on immune rejection of organ transplant

Immune rejection in organ transplantation was a headache problem. Although cell transplant rejection is much smaller than organ transplants, but the transplanted cells eventually running low with the passage of the time, in particular for the allogeneic transplant.

Now clinically use the chemical immunosuppressants usually, such as cyclosporin A and so on. However, these drugs have heavy side effects, such as lowering the systemic immune function, induced tumors and infection. At the same time also can damage the liver and kidney. Patients are often being forced to stop use because they can not tolerate the negative effects. So it is very meaningful to find an alternative immunosuppressants with lesser negative effects.

According to the modern studies, the immunomodulators in Chinese drugs are abounding. Some of they can extend the survival time of cardiac cells’ allogeneic transplants in mice, and can inhibit the immune rejection with lesser side effects. This shows that the promising of Chinese drugs on control immune rejection.

5.1. Effect of astragalus on CD4+CD25+ T cells and FOXP3/IL -10 in allografted mice

Objective: The experiments investigate the impact of high dose Astragalus on the survival period of the allograft, CD4+ CD25+ T cells, Foxp3 and some cytokines with murine skin transplantation model, at the same time compared with the traditional suppressive agent-CsA, so as to providing theoretical and experimental foundations of Astragalus for the therapy to allograft rejection and tolerance induction.

Methods: Established murine skin allo-transplantation model, administered different dose of Astragalus injection to know the most suitable dose through the effects on the survival time of allograft and mixed lymphocyte reaction (MLR). Experimental mice administered the effective dose of Astragalus injection and CsA in vivo. Survival condition of grafted skin and general condition of mice were observed daily. With 3H－TdR incorporation assay, we studied the effects of Astragalus injection and CsA on MLR. The levels of CD4+CD25+ T cells were examined by immunofluorescence staining and flow cytometry. Extracted total RNA from the spleen cells of mice, with RT-PCR and electrophoresis, we measured the expression of Foxp3 mRNA. Using radioimmunoassay (RIA), we examined the IL-10 and IL-2 concentration in murine serum and culture supernatant of spleen cells after administering Astragalus injection in allograft models.

Results: 1. Administering Astragalus injection (60g/kg) and combined with spleen cells injection in vivo after transplantation apparently prolonged the survival time of the allograft.

2. On the day 14 (the top period of the rejection), Astragalus injection (60g/kg) inhibited MLR, decreased specific memory response of T cells.

3. On the day 7 after transplantation, the CD4+CD25+ T cells level and the related expression of Foxp3 mRNA in groups of administering Astragalus injection (60g/kg)，combined with spleen cells injection and CsA increased and reached to the maximum on the day 14, then decreased and recovered the initial level on the day 21. The control group did not shown marked change as compared to preoperative level.

4. Compared with the control group, there was an obvious rise of group AST (60g/kg) and AST (60g/kg) + SP in the serum IL-10 on the day 1, 7, 14 post-transplantation by turns and a peak on the day 14 and then a descent afterwards; on the day 21, it dropped to initial level. Astragalus injection inhibited the IL-2 production on the day 14.

Conclusions: 1. Astragalus injection (60g/kg) and Astragalus combined with donor spleen cells previously injection can obviously prolong the survival period of the allografts in vivo.

2. Astragalus injection (60g/kg) and Astragalus combined with donor spleen cells previously injection in vivo can inhibit MLR in grafted mice.

3. After administering Astragalus injection (60g/kg) and Astragalus combined with donor spleen cells previously injection, CD4+ CD25+ T cells and its related molecules Foxp3/IL-10 have synchronous changes during the rejection of skin allograft and they all reached the top points in the peck of rejection.

4. Astragalus injection (60g/kg) or Astragalus combined with donor spleen cells previously injection can up-regulate the level of CD4+CD25+ T cells and its related molecules Foxp3/IL-10 in vivo.

5. Astragalus injection (60g/kg) can inhibit the IL-2 production in the process of allograft rejection.

5.2. The Effect of Chinese drugs on Acute Rejection in Renal Transplant Rats

Original Name of the article was <The Effect of Chinese Medicine AnTaiYangXue Mixture on Acute Rejection Reaction in Renal Transplantation Rats>. The AnTaiYangXue Mixture (ATYXM) is a compound preparation of Chinese drugs.

Objective: To study the effect of Chinese Medicine AnTaiYangXue Mixture (ATYXM) on acute rejection reaction of renal transplantation in rat and provide experimental basis for application of this recipe in organ transplantation.

Methods: Allogenic rat renal transplantation model was established from SD to Wistar and randomly divided into 4 groups, 12 rats in each group. Group 1:control group, nomal saline (NS) was administered intragastricly (10ml?kg-1?d-1)；Group 2: ATYXM group (10 ml?kg-1?d-1); Group 3：Cyclosporin A (CsA) group (10ml?kg-1?d-1); Group 4: Integrated traditional and western medicine group (ATYXM 10ml?kg-1?d-1+ CsA 2.5 ml?kg-1?d-1）.The recipient rat survival time was observed. Six rats were killed on the sixth day after operation to examine the histopathological changes of grafted kidney and to measure the levels of blood creatinine (Cre) and urea nitrogen (BUN).

Results: The rats of control group were all dead after 8 d and the average survival time was (6.50 ± 1.05) d. The average survival time of the rats in Group 2 was (8.50±1.05) d, group 3 was (9.00±1.41) d and group 4 was (12.33±2.42) d.

After 6 days of operation the transplanted renal tissues were sampled and examined. In group 1, the kidney rejection reaction was severe and the volume of transplanted renal increased clearly. It happened the interstitial edema and inflammatory cell infiltration, mainly lymphocytes diffuse infiltration, infiltration of the interstitial cells affected the renal tubular, some tubular was necrotic degeneration, demonstrated common inflammatory changes in the small tube. In group 1 and 2, the pathological manifestations was similar to the control group, the difference was not obvious under the microscope. However, the pathological changes were lighter than that of group 1. In group 4, glomerular vascular was slight expansion and oozing, a small number of infiltration of neutrophils, lymphocytes, mononuclear cell and a small number of interstitial cell infiltration. Showed the transplant renal acute rejection reaction was inhibited.

ATYXM and CsA could both prolong survival time of recipient rats. The combined application of CsA could markdedly prolong survival time of recipient rat. ATYXM coule also protect kidney function and improve pathological changes of transplanted kidney.

Conclusion: ATYXM has a definite inhibition effect on acute immune rejection reaction in renal transplantation, and might have a synergism with sub treatment dosage of Cyclosporin A (CyA).

5.3. Effects of Chinese drugs on survival time of allogeneic transplant of pig skin

Original Name of the article was <Effects of compound preparation of Cordyceps sinensis and Tripterygium hypoglaucum (CSTHC) on survival time of pig skin after allogeneic transplantation>.

Objective：To investigate the effects of compound preparation of Cordycepssinensis and Tripterygiumhypoglaucum on survival time of grafted pig skin after allogeneic transplantation and its mechanism.

Methods： Donor pigs, a healthy male, weight 78 kg. Recipient pigs: 17 Guizhou miniature white pigs, 3-month-old, were randomly divided into 3 groups: CSTHC group, cyclosporine A group and model group.

From donor pigs took 3 group of 4.0 cm × 4.0 cm graft, warm-up in each group’s incubation solution for 1h incubation, and then transplanted 9 pieces on the side of the chest and abdomen of each recipient pig. The 8th day after transplantation, using the corresponding ointment smeared the transplanted skin area once daily.

The survival time， the appearance and the histomorphological changes of the grafted pig skin were observed. The histomorphological changes of testicles in pigs were also examined. The CD4 and CD8 expressions in the grafted pig skins were measured by immunohistochemical method. The white blood cell count in peripheral blood and the liver and renal functions were also examined.

Survival standard of skin graft: Skin graft showed shiny red as survival. If it was dull pale or dark brown and dry area >2/3 then it was ecrosis.

Results：From the 9th day after transplantation, the skin graft of model group was starting progressive necrosis, the first 13th days was all necrosis. At this time, skin graft survival in good condition in CSTHC and CsA groups. The first 3 to 4 weeks after transplantation, the skin graft have emerged immune rejection one after another in CSTHC group and CsA group, respectively after the first 34 days and the first 35 days was all necrosis.

The survival time of the grafted pig skin in the CSTHC treated group was much longer than that of saline treated group. There was no remarkable difference in the survival time between the CsA treated group and the CSTHC treated group. The expressions of CD4 and CD8 were lower in the CSTHC treated group than those in the saline treated group on the 7th and 14th days after skin graft, while there was no significant difference in the indices between the CSTHC treated group and the CsA treated group. The WBC count was higher in the model group than those of in the CSTHC treated group or CsA treated group on the 7th day after skin graft.

Conclusion： CSTHC can prolong the survival time of allogeneic grafted pig skin. Its mechanism of inhibiting the immunological rejection may relate to decreasing the expressions of CD4+ and CD8+ in the grafted pig skin and reducing the local inflammatory reaction.