| Objective: Based on the research on the theory of the kidney dominating reproduction, to continue a study on pregnancy immunological regulating mechanism of a tonifying the kidney formula, Pill Zhu Yun â…¢, on treating threatened abortion or recurrent spontaneous abortion, also focus on the filial generation physical and intelligence development which completed maternal therapy in the hospital with herbal medicine for tonifying the kidney. On the other hand, to study the effect of the same formula on stimulating ovulation and regulating neuro-endocrine on kidney deficiency and ovulation inhibition rat model, as well as the effect on protecting fetus on a pregnant rat toxic model by taking fenpropathrin. To explore the further mechanism of Pill Zhu Yun â…¢ and expound the theory that the kidney being the origin of congenital constitution. Methods and Results:1. The research on theory of the kidney dominating reproductionTo collect and analyze the ancient and update traditional Chinese medical documents on the kidney functions and reproduction, tonifying kidney therapy on reproductive disorder, and research methods on kidney deficiency models and reproductive disorder models, and point out further research ideas.2. Studying clinical effect and mechanism of Pill Zhu Yun â…¢ on early pregnant women with threatened abortion30 patients were diagnosed as threatened abortion and the kidney deficiency or both kidney and spleen deficiency in the first trimester in the 1st Affiliated Hospital, GuangzhouUniversity of TCM. They were treated with Pill Zhu Yun III 6gm, 3 times per day for 4 weeks. Blood was' collected pre- and post-treatment for H-2 and IL-10 analysis by flow cytometry, as well as progesterone and P-HCG level by ELISA. 20 women in early pregnancy were measured the same items as the controlled group. Results: 26 patient were healed in the group. The effective rate is 86.67%. The IL-2 level was significantly higher in patients pre-treatment, compared with the controlled group (P <0.01) ,and the level was decreased post-treatment (P <0.01) The patients'IL-10 level was lower pre-treatment but not significantly compared with the controlled group (P =0.13>0.05 ) , and the level was increased significantly after the treatment (P <0.01) . Both EL-2 and IL-10 level were no significant different post-treatment compared with the controlled group (P >0.05) . The ratio of IL-2/IL-10 was significantly higher pre-treatment but normal post-treatment. The patients' progesterone and P-HCG level were lower pre-treatment (P <0.01, P <0.05) compared with the controlled group, and both increased significantly after the treatment (P <0.01) .3. A research on the filial generation physical and intelligence development which completed maternal therapy with herbal medicine for tonifying the kidney 100 children, aged 1-3, as the filial generation which completed maternal therapy with herbal medicine for tonifying the kidney were investigated for their physical and intelligence development. Their mothers were patients diagnosed as threatened abortion and the kidney deficiency or both kidney and spleen deficiency in the first trimester in the 1st Affiliated Hospital, Guangzhou University of TCM in the recent years. 50 children in the same age were investigated as a controlled group, which the mothers were normal in the whole pregnant period. The newborn babies' body length, weight, round length of head and chest were recorded. The present height, weight, round length of head, chest and arm were measured. The intellect's constitution were measured with Gesell developmental schedules. Results: The average age of the patients was 27 year-old, their children's age was 22.08-42.50 months(57 female and 43 male). The average age of the controlled group was 25, the children was 23.43-44.24 month-old (28 girls and 22 boys). All physical and intelligence items in the survey group were normal, without significantly lower comparedwith the controlled group. Both female and male newborn of the survey group had higher body length, the difference was significant compared with the controlled group (P<0.05) The survey girls' present weight and height were higher than that of girls in the controlled group (P<0.05) . Language ability of the survey girls and boys, reaction abilities to human and material of the survey boys was better than that of the controlled group (P<0.05 ) . 4. Study on the effect of Pill Zhu Yun III on stimulating ovulation and regulating neuro-endocrine on kidney deficiency and ovulation inhibition rat model SPF grace female SD rats, aged 10 weeks, weight 200-250gm, were fed with hydroxyurea 300mg/Kg daily for creating a kidney deficiency and ovulation inhibition model. The rats were divided into 5 groups: A: model without herbs, B: model with high dose herbal medicine, C: model with middle dose herbal medicine, D: model with low dose herbal medicine, E: controlled group(fed with saline instead of medicine). There were 10 days for creating model and treatment. Results: Group A, B, C and D showed some symptoms of kidney deficiency, such as withered fur, slow action and less ability. The body weight of rats in each group were all increased from Dl to D10, the different was no significant, P>0.05. The weight of pituitary, adrenal gland, kidney and spleen in each group were similar, the different between each 2 groups was no significant, P>0.05. But the weight of ovary and uterus of group A were the lowest, there was significantly different compared with that of group E (P<0.05) Group C's weight ratio of ovary/body and uterus/body were significantly higher than that of group A, P<0. 05. Group D's uterus weight and the weight ratio of uterus/body were significantly higher than that of group A, /><0. 05 and P<0. 01. Sera P -EP level in each groups was not significantly different on DL.But on D10, the level of the animals in group A and D was significantly lower than that in group B, C and E (PO.05, PO.01) . It is suggested that the high and middle dose of Zhu Yun EH could resist ï¿¡ -EP inhibition caused by hydroxyurea in this kidney deficiency model. Sera FSH in group B and C on D10 was much higher than that on Dl (PO.05) . E2 level in each groups on Dl and 10 was not much different. Testosterone level in group A and E on D10 was not much different, but in group B the level was higher than that on Dl (/><0.05). Sera cortisol level in group A on D10 was significantly lower than that on Dl(P<0. 05), and alsolower than that in group E(P<0. 05). Cortisol level in group B, C, D and E on Dl and D10 was not much different. Ovary FSH receptor (FSH-R) in group B and E had the strongest expression. FSH-R in group A was weaker than that in group B and E (P<0.05). Estrogen receptor(ER) in endometrium was expressed stronger in group B, C and D, but significantly weaker in group A, P <0.01. ER expression also had significantly different between group E and A, P <0.05. The numbers of primary follicles in ovary of each group were not much different, but the numbers of mature follicles in group B and E were much more than that in group A (P<0.05). The endometra of group A and D was thinner, but in group B and C, the endometra was thicker than group A, the gland was in late proliferative phase or early luteal phase. The endometra of group E was normal. 5. Study on the effect of Pill Zhu Yun HI on resisting reproductive toxic and feto-neuro-toxicity to pregnant rat model toxified with fenpropathrin SPF grace female SD rats, aged 10 weeks, weight 200-250gm, fed with fenpropathrin on the first day (Dl) of conception. The dose of fenpropathrin was 1/20 of its LD50. Groups division: A: pure fenpropathrin model; B: 20% fenpropathrin model; C: pure fenpropathrin model with high dose herbal medicine; D: pure fenpropathrin model with middle dose herbal medicine; E: pure fenpropathrin model with low dose herbal medicine; F: pure fenpropathrin model with taurine; G: normal pregnant control. There were 15 days for creating model and treatment. Results: The rats fed with fenpropathrin didn't show any toxic symptoms and signs. The body weight of the animals was increased after conception. The difference between each two group was not significant. The rats in group A present a highest aborting rate as 54.54%, significantly higher than all other groups* P<0.01. The average fetus number and alive fetus rate in group A were lower, compared with other groups, P<0.01. The fetus absorbed rate in group A was 30. 71%, higher significantly than group B, D and G, P<0.05. Cx43 mRNA expression in fetus rat tissues was high in group B, it was significantly higher than group E C^O.05) . Pax3 mRNA expression in fetus tissues also was high in group B, but no significant difference compared with other groups. Cx43 protein in fetus brain was stronger in group A and B, but weaker in group D and F. compared group A with D and F, P<0.05.; Group B compared with F> P-0.01. The bodyweight of fetus in each group was similar, the different was not significant. The length of fetus body in group A was shorter than group D, F and G, P<0.05. The pregnant rats' sera E2 level was highest on D8, compared with group C, D and F, P<0.05. On D8, sera progesterone level was higher in group B and C, compared with group G. P>0.05. Sera IFN-y level of group A on D8 was significantly higher than that of group C and G, P<0.05, P<0.01. On D15, the ratio of IFN-y/ IL-4 of group A was higher than that of group G, P<0.05. It is suggested that the balance of TH1/TH2 cytokine in group A is inclined to the side of TH1. The weight of uterus and the ratio of uterus/body of group A was the lowest compared with that of group C, D and E, P<0.05. The weight of liver and spleen of group A also was the lowest, but only compared with that of group F it was significant, P<0,05, Conclusion:1. The theory of the kidney dominating reproduction was formed in a long history period and developed in recent decays on the research of clinical works and basic experiments.2. The tonify-kidney formula Zhu Yun III showed its curative effect on threatened abortion with syndromes of kidney deficiency or spleen-kidney deficiency. The balance of cytokine TH1/TH2 was inclined to the side of TH2 and the level of progesterone and ï¿¡ -HCG were increased after the treatment.3. The filial generation physical and intelligence development which completed maternal therapy with herbal medicine for tonifying the kidney was normal compared with same age children without any maternal treatment during conceptive period. It is safe for filial generation while maternal therapy in tonify-kidney and calming fetus with herbal medicine.4. In experimental research, Formula Zhu Yun HI showed its effects on regulating neuro-endocrine functions to improve follicle develop and mature, increase the expression of FSH-R and ER in kidney deficiency and ovulation inhibition rat model.5. In experimental research, Formula Zhu Yun III also showed its effects on maternal-fetus protection in pregnant rat model toxified with fenpropathrin. On the sensitive period to teratogenic agent, the herbal formula may insist the reproductive toxic and...
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