Regulatory Effect of Liuwei Dihuangtang and Guifu Dihuangtang on Low-dose Hydrocortisone Reduction and Discontinuance in Mice with Drug-induced Deficiency Syndrome / 中国实验方剂学杂志

Objective:To study the effect of the kidney-tonifying recipes on mouse syndrome and adrenal function induced by low-dose hydrocortisone reduction and discontinuation. Method:The 48 male ICR mice were randomly divided into normal control group, hydrocortisone group, Liuwei Dihuangtang group and Guifu Dihuangtang group, with 12 rats in each group. Each group was given 1.32 mg·kg-1·d-1 hydrocortisone for 28 days, and half a dose for 7 days, and the administration was discontinued 14 days before modeling. In the reduction and discontinuation phase, 12.5 g·kg-1·d-1 Liuwei Dihuangtang and 13.5 g·kg-1·d-1 Guifu Dihuangtang were simultaneously administered to the corresponding group for 21 d. On the 28th and 49th day of the experiment, the characteristic information of mice was detected by the experimental methodology of syndrome differentiation and treatment in mice (holding power, body surface infrared temperature). The mice were sacrificed on the 50th day of the experiment, the spleen and thymus were separated, and the spleen and thymus index were calculated, serum corticosterone content is measured by enzyme-linked immunosorbent assay (ELISA), the total RNA of the adrenal gland was extracted, and the expressions of Star, cholesterol-side-chain cleavage enzyme (Cyp11a1), Cyp21a1, Cyp11b1, low-density lipoprotein receptor (Ldlr), scavenger receptor class B type I (Scarb1/SRB1), Hmgcr, acyl-CoA-cholesterol acyltransferase-1 (Acat1), hormone sensitive lipase (Lipe/HSL), insulin-inducible gene 1 (Insig1) and sterol regulatory element binding transcription factor 2 (Srebf2) were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Western blot was used to detect the expressions of LDLR, SRB1, CYP11A1 and cytochrome P450 family 21 subfamily A member 2 (CYP21A2) in adrenal gland. Result:Compared with the normal control group, the body weight and the body mean infrared temperature of the model group were significantly decreased after 28 days of continuous administration (P<0.01). After the administration of the drug, the mice in the model group showed a significant decrease in the grip strength (P<0.05), a significant increase in the thymus index (P<0.01), a significant decrease in the serum corticosterone content (P<0.05). Expressions of adrenal Cyp21a1 and Hmgcr genes decreased (P<0.05, P<0.01), Lipe and Acat1 gene expressions increased significantly (P<0.01), adrenal CYP11A1, SRBI protein expressions decreased significantly, and LDLR protein expression increased. Compared with the model group, the body weight of Guifu Dihuangtang group decreased significantly after 21 days of treatment with traditional Chinese medicine (P<0.01), the spleen index of Liuwei Dihuangtang group decreased significantly (P<0.01), expressions of Cyp11a1, Cyp21a1, Acat1 and Lipe genes in Guifu Dihuangtang group were significantly increased (P<0.05, P<0.01), expressions of Ldlr and Scarb1 genes were significantly decreased (P<0.05), and expressions of Ldlr, Acat1 and Lipe genes in Liuwei Dihuangtang group were significantly decreased (P<0.05, P<0.01), and expressions of CYP11A1 and SRBI proteins in Liuwei Dihuangtang and Guifu Dihuangtang increased. Conclusion:Guifu Dihuangtang can promote the recovery of adrenal function in mice with drug-induced deficiency syndrome by correcting expression of adrenal cortex synthase.

Research Progress in PTEN as Target Spot in TCM for Prevention and Treatment of Primary Liver Cancer / 中国中医药信息杂志

Phosphatase and tensin homolog deleted on chromosome ten (PTEN) is an important negative regulatory molecule in the development and progression of primary liver cancer and other cancers. TCM has unique advantages in the prevention and treatment of primary liver cancer. This article reviewed the research progress in PTEN as target spot in TCM for prevention and treatment of primary liver cancer in the past 10 years, and provided references for further research and clinical application.

Effect of Osthole on Adrenocortical Function in Y1 Mouse Adrenocortical Tumor Cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.</p><p><b>METHODS</b>Y1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).</p><p><b>RESULTS</b>Y1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.</p><p><b>CONCLUSION</b>Ost could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.</p>

The correlation between adrenal corticosteroids and cytokines expressions in mice of different syndromes / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the cytokines expressions in the adrenal gland and its correlation with serum adrenal corticosteroids in mice of different syndromes.</p><p><b>METHODS</b>Using the quantitative four diagnosis and syndrome differentiation methods, 60 normal mice and 190 H22 liver cancer bearing mice were syndrome typed. Serum corticosterone and aldosterone were tested by ELISA, and mRNA expressions of cytokines in the adrenal gland were detected using Real-time PCR.</p><p><b>RESULTS</b>Mice of different syndromes were obtained, such as normal mice of no syndrome, normal mice of vigorous qi syndrome, normal mice of qi deficiency syndrome, liver cancer bearing mice of excessive evil toxic syndrome, liver cancer bearing mice of evil lying in the middle syndrome, liver cancer bearing mice of weak evil toxic syndrome, and liver cancer bearing mice of poisonous pathogenic factors and qi deficiency syndrome. The serum corticosteroids were significantly higher in the liver cancer bearing mice than in the normal mice (P < 0.05). The cortex hormones increased most significantly in the liver cancer bearing mice of excessive evil toxic syndrome (P < 0.05). Compared with the normal mice, IL-1beta, IL-2, IL-6, IL-10, IL-12alpha, IL-12beta, and TNF-alpha gene expressions increased in the liver cancer bearing mice, while only expressions of IL-1alpha and IL-5 decreased. But the expressions of IL-13 and transforming growth factor beta1 (TGF-beta1) showed no regularity. The expressions of IL-4 and INF-alpha were not detected in all mice. It is notable that the more severe degree of poisonous pathogenic factors, the higher the expressions of serum corticosterone and aldosterone levels as well as IL-6, the lower expressions of IL-1beta, IL-2, IL-5, IL-12alpha, IL-12beta, and TNF-alpha.</p><p><b>CONCLUSIONS</b>The increased serum corticosteroid level in liver cancer bearing mice could possibly be induced by chronic tumor stress, partial cytokines were involved in the synthesis and secretion of the adrenal hormone. Of them, IL-6 might positively regulate the secretion of corticosteroids, while IL-1beta, IL-2, IL-5, IL-12alpha, IL-12beta, and TNF-alpha might negatively regulate their secretions.</p>

Analysis of the dynamic changes of blood hormone levels in H22 liver cancer mice of poisonous pathogenic factors syndromes to different degrees / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the dynamic changes of blood hormone levels in H22 liver cancer mice of poisonous pathogenic factors syndromes (PPFS) to different degrees.</p><p><b>METHODS</b>Two hundred and twenty mice were injected with H22 tumor cells from their armpits. On the ninth day after inoculation the mice of severe poisonous pathogenic factors syndrome (SPPFS) and of mild poisonous pathogenic factors syndrome (MPPFS) were screened. Besides, another normal control group consisting of 30 mice was set up. The mice were killed on the tenth and eleventh day after inoculation (as the 1st and 2nd time window). The weight of the tumor, the wet weight of the thymus and the spleen were weighed. The plasma adrenocorticotropic hormone (ACTH), corticosterone, aldosterone, thyroid hormone T3 and T4, testosterone, TNF-alpha, and IFN-gamma were detected by ELISA. All the aforesaid laboratory parameters were analyzed.</p><p><b>RESULTS</b>The tumor weight was obviously larger in mice of the SPPFS group than in those of the MPPFS group at the same time window (P < or = 0.05). Compared with the normal control group, the thymus was obviously atrophied (P < or = 0.05), the spleen was significantly enlarged (P < or = 0.05), the plasma ACTH significantly increased (P < or = 0.05) in the SPPFS group at the two time windows. But the increment of ACTH was less in the MPPFS group. The plasma corticosterone showed similar tendency as that of ACTH. At the 1st time window the plasma testosterone significantly increased in the two groups (P < or = 0.05). The plasma testosterone and T4 showed a decreasing tendency in the SPPFS group. The plasma TNF-alpha and IFN-gamma levels showed an increasing trend in the two groups. Correlation study showed that the degree of PPFS was negatively correlated with qi deficiency (r = -0.766, P < or = 0.05) and T4 (r = -0.738, P < or = 0.05). The degrees of PPFS was positively correlated with the plasma ACTH level (r = 0.635, P < or = 0.05). The degree of qi deficiency was positively correlated with yang heat syndrome (r = 0.632, P < or = 0.05). The plasma ACTH was negatively correlated with T4 (r = -0.504, P < or = 0.05). The plasma testosterone was positively correlated with TNF-alpha (r = 0.619, P < or = 0.05).</p><p><b>CONCLUSIONS</b>PPFS occurs naturally and shows difference to different degrees in the development of H22 liver cancer. The disorders of neuroendocrine hormones and the suppression of the immune function show dynamic changing trends.</p>

Characteristics of gene expression of adrenal cortical steroid synthetase and its regulatory factor in mice with H22 liver cancer of different patterns / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the characteristics of gene expression of adrenal cortical steroid synthetase and its regulatory factor in mice with H22 liver cancer of different patterns.</p><p><b>METHODS</b>Syndromes revealed in mice with H22 tumor were differentiated by quantified four diagnostic methods and syndrome differentiation, and mice with commonly encountered patterns (A: evil-toxin accumulation pattern, B: qi-deficiency pattern, C: yang-qi deficiency pattern and D: qi-yin-yang deficiency pattern) were screened out for subjecting to the study. Two batches of GeneChip Mouse Exon 1.0 ST Array detection were performed in the selected mice for detecting the gene expressions of adrenal cortical steroid synthetase and its regulatory factor, with the analysis performed put stress on the differential expressions in mice of various syndrome patterns.</p><p><b>RESULTS</b>Data obtained from the two batches detection showed well repeatability, in which similar genes of high or low expression emerged. The adrenal cortical steroid synthetase genes, such as Cyp11a1, Star, Cyp11b2, Cyp21a1, Hsd3b and Hsd17b were highly expressed, with few difference among the four patterns. However, Cyp11a1 was down-regulated and Cyp1b2 up-regulated in all patterns; Hsd3b1 and Cyp21a1 down-regulated in pattern A and B, but up-regulated in pattern C and D. As for the expressions of the relative regulatory factors, Cyb5b and Wnt4 were down-regulated but Fdx1, Fdxr, Hsd11b1, Por, Agt and Nr 0b1 were up-regulated in all patterns; Nr5al down-regulated in pattern A but up-regulated in other three patterns; Nr4al and Nr4a2 up-regulated in pattern A and down-regulated in the others.</p><p><b>CONCLUSIONS</b>The adrenal cortical steroid synthetase genes are rather conservative and stable in mice bearing H22 liver cancer, part of the expression might be correlated to the condition of disease and essence of syndromes, embodying the differences among different patterns in the same disease.</p>

Discussion on some basic academic problems in research and development of treatment based on syndrome differentiation / 中国中西医结合杂志

To implement treatment depending on syndrome differentiation, namely, the syndrome dependent therapy (SDT), is the characteristics of Chinese medicine, also an important field for TCM researches and development in the contemporary era. This paper summarized the common academic problems in this field, involving the relationship between syndrome and disease, body constitution, four diagnostic methods and SDT, the mechanisms of diseases, syndromes and SDT, as well as the relationship between SDT and Western medicine. The thinking path and recommendation for solution were proposed as well.

Effect of er xian decoction and its subdivisions on granulosa cells secretory function in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effects of Er Xian Decoction (EXD) and its three subdivisions "warming Shen" ,"nourshing Yin" and "adjusting Chong and Ren" in regulating the level of estradiol (E2) on the primary cultured granulosa cells.</p><p><b>METHODS</b>Effect of EXD and its three subdivisions, also part of the effective components of this formula, icariin and curculigoside, on level of E2 were carried out using primary cultural granulose cell as the experimental model.</p><p><b>RESULT</b>EXD and its three subdivisions could stimulate the secretion of E2, especially the "warming Shen" group (P <0.05). All the composing of Chinese herbs of this formula could promote the level of E2 in different degree, and the Rhizoma Curculiginis, Radix Moromade Officinalis, and Herba Epimedii have the best effects (P <0. 01).</p><p><b>CONCLUSION</b>The regulation of EXD on the hypothalamus-pituitary-gonad (HPG) axis may be related to promoting the secreting of E2 at the site of granulosa cell. The "warming Shen" subdivision has the better effect in promoting the secretion of E2.</p>