Twelve-component pharmacokinetic study of rat plasma after oral administration of You-Gui-Wan in osteoporosis rats with kidney-yin deficiency and kidney-yang deficiency.

You-Gui-Wan is a widely used traditional Chinese medicine preparation for the treatment of osteoporosis with kidney-yang deficiency, and is composed of both yang-invigorating and kidney-tonifying herbs, and yin-nourishing and kidney essence-replenishing herbs. Considering that the pharmacokinetics of drugs might differ in different pathological conditions, it is necessary to study the pharmacokinetic characteristics of You-Gui-Wan under different osteoporotic conditions. In this study, the pharmacokinetic behaviors of You-Gui-Wan in osteoporosis rats with kidney-yin and kidney-yang deficiency were compared. The results showed that the absorption, metabolism, and disposition of You-Gui-Wan varied widely in animals with different types of osteoporosis. The active components belonging to the yang-invigorating herbs, such as aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypacoitine, benzoylmesaconine, chlorogenic acid and pinoresinol diglucoside, had a higher uptake and slower elimination in osteoporosis rats with kidney-yang deficiency, which corresponds to the opinion that You-Gui-Wan is used to treat kidney-yang deficiency syndrome, and indicates the scientific nature of Bian-Zheng-Lun-Zhi.

The Effect of Shen Qi Wan Medicated Serum on NRK-52E Cells Proliferation and Migration by Targeting Aquaporin 1 (AQP1).

BACKGROUND Shen Qi Wan (SQW) as a well-known formula for the amelioration of kidney yang deficiency syndrome (KYDS), and it has been widely employed in traditional Chinese medicine (TCM). This study aimed to investigate the effect and underlying mechanism of SQW medicated serum on proliferation and migration in NRK-52E cells. MATERIAL AND METHODS We employed the real-time cell analysis (RTCA) system to investigate the effect of SQW medicated serum on proliferation and migration in NRK-52E cells. In addition, the migration was further investigated by using a wound-healing assay. The mRNA and protein expression level of aquaporin 1 (AQP1) of NRK-52E cells with SQW medicated serum-treated were quantified by real-time quantitative polymerase chain reaction (q-PCR) and western blot assay, respectively. Furthermore, NRK-52E cells were transfected with lentivirus AQP1-RNAi to assess migratory cell abilities in vitro. RESULTS The migratory abilities of NRK-52E cells were significantly increased after SQW medicated serum treatment (P<0.05), and no significant difference in cell proliferation. In addition, SQW medicated serum was significantly upregulated the mRNA and protein expression level of AQP1 in NRK-52E cells (P<0.05). Additionally, the in vitro metastasis test proved that knockdown of AQP1 suppressed migratory abilities according to RTCA and wound healing test while was reversed by SQW medicated serum (P<0.05). CONCLUSIONS Our study demonstrates that SQW medicated serum effectively promotes the migration of NRK-52E cells by increasing AQP1 expression, and AQP1 may be as a therapeutic target of SQW for renal injury treatment under KYDS.

Er Shen Wan extract alleviates polyuria and regulates AQP 2 and AVPR 2 in a rat model of spleen-kidney Yang deficiency-induced diarrhea.

ETHNOPHARMACOLOGICAL RELEVANCE: Er Shen Wan (ESW), has been empirically used for treating spleen-kidney Yang deficiency (SKYD) syndrome in Traditional Chinese medicine (TCM) for centuries and shows a variety of activities. The medicinal formula is a mixture of two component herbs, Psoraleae Fructus (PF, Bu-Gu-Zhi in Chinese) and Myristicae Semen (MS, Rou-Dou-Kou in Chinese). The current study was designed to evaluate ESWP antidiuretic treatment of polyuria and to explore potential mechanisms of renal water metabolism in the rat model of SKYD-induced diarrhea. MATERIALS AND METHODS: An animal model of 'SKYD-induced diarrhea syndrome' has been established to evaluate the therapeutic effect and action mechanism according to the clinical syndrome and symptoms. The optimal dose (3.5 g/kg) of ESWP was given to rats by gavage for two weeks. Urinary volumes after 24 h were recorded. After the end of the trial, macroscopic morphological and histological examination of the kidney were conducted. Serum levels of Arginine vasopressin (AVP) and aldosterone (ALD) were also measured. Additionally, quantitative real-time RT-PCR (RT-qPCR) and immunohistochemistry (IHC) analyses were performed to clarify the regulation of aquaporin 2 (AQP 2) and arginine vasopressin type 2 receptor (AVPR 2) in the kidney at the gene and tissue expression levels respectively. RESULTS: After the administration of ESWP, urinary output volume after 24 h was found to be significantly decreased in rats. Elevated plasma levels of AVP and ALD were detected. Histological kidney damage appeared to be impeded, and histological disease scores were reduced. In addition, the expression levels of AQP 2 and AVPR 2 were significantly increased. CONCLUSION: This study suggests that ESWP may elicit significant effects on the treatment of polyuria. Potential mechanisms at least partially involve hormone regulation, and alleviating renal pathological damage. Simultaneously, ESWP may alter renal water absorption by increasing AQP 2 and AVPR 2 expression levels. Thus, the in vivo experimental evidence indicates that ESWP has a therapeutic effect on the SKYD syndrome, which is consistent with its traditional usage.

miRNA Expression Profile of Saliva in Subjects of Yang Deficiency Constitution and Yin Deficiency Constitution.

BACKGROUND/AIMS: Based on the theory of constitution in Traditional Chinese Medicine (TCM), the Chinese Han population has been classified into nine constitutions. Of these, Yang deficiency constitution mainly exhibit cold intolerance while Yin deficiency constitution mainly exhibit heat intolerance. Some studies have been carried out to explore the modern genetic and biological basis of such constitution classification, but more remains to be done. MicroRNA (miRNA) serves as post-transcriptional regulators of gene expression and may play a role in the classification process. Here, we examined miRNA expression profile of saliva to further improve the comprehensiveness of constitution classification. METHODS: Saliva was collected from Chinese Han individuals with Yang deficiency, Yin deficiency and Balanced constitutions (n=5 each), and miRNA expression profile was determined using the Human miRNA OneArray®v7. Based on 1.5 Fold change, means log2|Ratio|≥0.585 and P-value< 0.05, differentially expressed miRNA was screened. Target genes were predicted using DIANA-TarBasev7.0 and analysis of KEGG pathway was carried out using DIANA-mirPathv.3. RESULTS: We found that 81 and 98 differentially expressed miRNAs were screened in Yang deficiency and Yin deficiency constitution, respectively. Among them, 16 miRNAs were identical and the others were unique. In addition, the target genes that are regulated by the unique miRNAs were significantly enriched in 27 and 20 signaling pathways in Yang deficiency and Yin deficiency constitution, respectively. Thyroid hormone signaling pathway is present in both constitutions. These unique miRNAs that regulated target genes of thyroid hormone signaling pathway may be associated with cold intolerance or heat intolerance. CONCLUSION: The results of our study show that Yang deficiency and Yin deficiency constitutions exhibit systematic differences in miRNA expression profile. Moreover, the distinct characteristics of TCM constitution may be explained, in part, by differentially expressed miRNAs.

Er Shen Wan extract reduces diarrhea and regulates AQP 4 and NHE 3 in a rat model of spleen-kidney Yang deficiency-induced diarrhea.

INTRODUCTION: Er Shen Wan (ESW), a traditional Chinese medicinal formula comprised of Psoraleae Fructus (Babchi seeds, from Psoralea corylifolia Linn.) and Myristicae Semen (Nutmeg, from Myristica fragrans Houtt.), is widely used to treat spleen-kidney Yang deficiency (SKYD)-induced diarrhea. Previous studies have demonstrated preliminarily that the petroleum ether extract of ESW (ESWP) exhibits significant anti-diarrheal activity. The present study aimed to evaluate the anti-diarrhea activity of ESWP and to explore the underlying mechanisms with respect to fluid metabolism in a rat model of SKYD-induced diarrhea. MATERIALS AND METHODS: A high-performance liquid chromatography-diode array detector (HPLC-DAD) approach was developed and validated for qualitative and quantitative analyses of the main constituents of ESWP. SKYD model rats were established and treated with an effective dose (3.5?g/kg) of the extract for two weeks. Anti-diarrheal activity and stool properties were observed. After the experiment, the appearance and histology of the intestines were evaluated. Serum levels of neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) were also determined. Furthermore, to characterize the regulation of aquaporin-4 (AQP 4) and Na+/H+ exchanger isoform 3 (NHE 3) in the colon, quantitative real-time RT-PCR (qRT-PCR), immunohistochemistry (IHC) and Western blotting (WB) were employed to detect mRNA and protein expression levels. RESULTS: In the rat models, oral ESWP administration significantly reduced the diarrhea score and the number and weight of wet stools. Jejunal and ileac histological damage was impeded, and the histology score decreased. Serum VIP levels were significantly decreased, in contrast to NPY levels. In addition, AQP 4 and NHE 3 expression levels increased significantly. CONCLUSIONS: These results showed that ESWP's anti-diarrheal effect might at least partially involve the regulation of hormones intimately involved in maintaining fluid and electrolyte levels, as well as by increasing AQP 4 and NHE 3 expression levels and enhancing the absorption of Na+ and water.

An integrated chinmedomics strategy for discovery of effective constituents from traditional herbal medicine.

Traditional natural product discovery affords no information about compound structure or pharmacological activities until late in the discovery process, and leads to low probabilities of finding compounds with unique biological properties. By integrating serum pharmacochemistry-based screening with high-resolution metabolomics analysis, we have developed a new platform, termed chinmedomics which is capable of directly discovering the bioactive constituents. In this work, the focus is on ShenQiWan (SQW) treatment of ShenYangXu (SYX, kidney-yang deficiency syndrome) as a case study, as determined by chinmedomics. With serum pharmacochemistry, a total of 34 peaks were tentatively characterised in vivo, 24 of which were parent components and 10 metabolites were detected. The metabolic profiling and potential biomarkers of SYX were also investigated and 23 differential metabolites were found. 20 highly correlated components were screened by the plotting of correlation between marker metabolites and serum constituents and considered as the main active components of SQW. These compounds are imported into a database to predict the action targets: 14 importantly potential targets were found and related to aldosterone-regulated sodium reabsorption and adrenergic signaling pathways. Our study showed that integrated chinmedomics is a powerful strategy for discovery and screening of effective constituents from herbal medicines.

Phenotypic characterization of nanshi oral liquid alters metabolic signatures during disease prevention.

This paper was designed to investigate the phenotypic characterization of Nanshi Oral Liquid (NOL) alters metabolic signatures of the 'Kidney Yang Deficiency syndrome' (KYDS). Urine metabolites were profiled by UPLC-ESI-Q-TOF-HDMS. The significantly changed metabolites such as xanthurenic acid, 4,8-dihydroxyquinoline, 3-methyldioxyindole, 4,6-dihydroxyquinoline, kynurenic acid, hippuric acid, taurine, tyramine, and 3-metanephrine, had been identified, and were related to the disturbance in tyrosine metabolism, steroid hormone biosynthesis, taurine and hypotaurine metabolism, tryptophan metabolism, phenylalanine metabolism and lysine degradation, which were helpful to further understanding the KYDS and intervention mechanism of NOL. The biochemical result showed that NOL can alleviate the kidney impairment induced by KYDS. Metabolomics results indicated the significantly changed metabolites were found to be reasonable in explaining the action mechanism of NOL. Interestingly, the effectiveness of NOL against KYDS was proved using the established metabolomics method and regulated the biomarkers as well as adjusted the metabolic disorder pathways. NOL had potentially pharmacological effect through regulating multiple perturbed pathways to normal state. This work showed that the metabolomics method was a powerful approach for studying the phenotypic characterization of disease's syndrome during disease prevention and its intervention mechanism.

[Regulatory mechanism of hormones of the pituitary-target gland axes in kidney-Yang deficiency based on a support vector machine model].

OBJECTIVE: To study the development mechanism of kidney-Yang deficiency through the establishment of support vector machine models of relevant hormones of the pituitary-target gland axes in rats with kidney-Yang deficiency syndrome. METHODS: The kidney-Yang deficiency rat model was created by intramuscular injection of hydrocortisone, and contents of the hormones of the pituitary-thyroid axis: thyroid stimulating hormone (TSH), 3,3',5-triiodothyronine (T3) and thyroxine (T4); hormones of the pituitary-adrenal gland axis: adrenocorticotropic hormone (ACTH) and cortisol (CORT); and hormones of the pituitary-gonadal axis: luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (T), were determined in the early, middle, and advanced stages. Ten support vector regression (SVR) models of the hormones were established to analyze the mutual relationships among the hormones of the three axes. RESULTS: The feedback control action of the pituitary-adrenal axis began to lose efficacy from the middle stage of kidney-Yang deficiency. The contents all hormones of the three pituitary-target gland axes decreased in the advanced stage. Relative errors of the jackknife test of the SVR models all were less than 10%. CONCLUSION: Imbalances in mutual regulation among the hormones of the pituitary-target gland axes, especially loss of effectiveness of the pituitary-adrenal axis, is one pathogenesis of kidney-Yang deficiency. The SVR model can accurately reflect the complicated non-linear relationships among pituitary-target gland axes in rats with of kidney-Yang deficiency.

The Fourier transform infrared spectra of the key organs derived from Kidney (Shen)-yang deficiency syndrome mice.

OBJECTIVE: To explore an approach to rapidly and accurately identify the compounds as biomarkers of Chinese medicine (CM) syndromes. METHOD: The Fourier transform infrared (FT-IR) spectrometry was applied to investigate the characteristic components of a mice model of Kidney (Shen)-yang deficiency syndrome (KDS), and the remedial effect of a typical CM formula Shenqi Pill (). Thirty-six females and 18 males of Balb/c mice were randomly divided into KDS, Shenqi or control group. The females and males of the same group freely were mated for 96 h, and the males were taken out and only the female mice were raised. Females of the KDS group were threatened by a ferocious cat every other day for 14 d. After delivery, the KDS, or gestational threatened, offspring were raised at standard condition for 11 weeks. Then 10 male offspring were randomly selected, anaesthetized and their representative organs, i.e. testes, kidneys, lungs and feet were collected, for the FT-IR scan. Mice of the Shenqi group were intragastric administered Shenqi Pill; while mice in the KDS and control groups were given the same volume of saline. RESULTS: The attenuated birth outcomes of the KDS group were displayed. The remarkable FT-IR differences of all organs between KDS mice and healthy control were mainly at 1,735-1,745 cm(-1) (indicating the increased levels of lipids) and at 1,640-1,647 cm(-1) and 1,539-1,544 cm(-1) (displaying the decreased proteins). No statistic FT-IR difference between Shenqi and control mice was observed. CONCLUSION: In accordance with major traits of KDS, prenatal stress extensively impaired the building up of proteins and resulting in the excessive lipid storage, and FT-IR could effectively identify the biomarkers of KDS.

[Research on building method of spleen kidney Yang deficiency diarrhea rats model].

Spleen kidney Yang deficiency (SKYD) diarrhea is a common syndrome in tranditional Chinese medicine (TCM). Until now, there is not an ideal SKYD diarrhea rat model for the research. In this study, we compared single factor way (method I, injecting hydrocortisone and gavaging Sennae Folium) with compound factors way(method II, gavaging adenine, improper diet, exhaustion, and gavaging Sennae Folium) on establishing SKYD diarrhea rat model. After modelling, diarrhea index, D-xylose excretory rate, NOS/cGMP signal transduction system, organ index and histopathology examination were used to evaluate the two ways. The results showed that, compared with health group, all the assessment criterias of method I and method II had significant differences (P < 0.01, 0.05). In addition, the index such as diarrhea index, NOS/cGMP signal transduction system, organ index (kidney, testis and thymus) and histopathology examination had significant differences (P < 0.01, 0.05) between method I and method II. In conclusion, the compound factors modelling method better conforms to the symptom of diarrhoea model caused by SKYD. This new modelling method provides a basis for studying on TCM astringents warming and tonifying the spleen and kidney, relieving diarrhea.

Bone marrow angiogenesis in patients presenting with differential Chinese medicine syndrome: correlation with the clinico-pathological features of aplastic anemia.

OBJECTIVE: To explore differences in bone marrow angiogenesis seen in aplastic anemia (AA) patients presenting with differential Chinese medicine (CM) syndrome, and to correlate these differences with clinical pathology. METHODS: Thirty-five patients were enrolled, including 18 with "yang deficiency syndrome" and 17 with "yin deficiency syndrome." Bone marrow biopsies and serum were collected. Microvessel density (MVD) and positive expression of vascular endothelial-derived growth factor (VEGF) were detected by immunohistochemisty. Hypoxia inducible factor -1α (HIF-1α), and VEGF expression were assayed by enzyme-linked immunoabsorbent assay (ELISA), serum lactate dehydrogenase (LDH) was tested by enzyme method and liquid chip technology was used to detected the expression of interleukin (IL)-2, IL-4, IL-6, IL-10, interferon (IFN)-γ and tumor necrosis factor (TNF)-α. RESULTS: Counts for leukocytes, absolute neutrophils and platelets in "yin deficiency syndrome" were lower than those found in "yang deficiency syndrome" (P<0.05). MVD and VEGF expression, and the positive rate of CD34 and VEGF in bone marrow were lower in AA, especially in "yin deficiency syndrome" (P<0.01 or P<0.05). "Yin deficiency syndrome" displayed decreased VEGF and LDH expression, and enhanced expression of HIF-1α as compared to "yang deficiency syndrome" (P<0.05). Levels of IL-4 and IL-6 were higher in AA (P<0.01), but IL-10 was decreased (P<0.05). High TNF-α expression was seen in "yang deficiency syndrome" and IFN-γ expression was decreased in "yin deficiency syndrome" as compared with normals (P <0.01 and P<0.05, respectively). CONCLUSION: AA patients have lower MVD than normals, especially in "yin deficiency syndrome." MVD might differentially correlate to disease severity, and could be dependent on bone marrow or serum VEGF expression and LDH. Additionally, IL-2, IL-10, IL-4 and IFN-γ were negatively associated while IL-6 and TNF-α were positively associated with MVD.

[Treatment of ulcerative colitis with spleen and kidney yang deficiency by kuijiening plaster: a randomized controlled study].

OBJECTIVE: To explore the clinical efficacy of ulcerative colitis with spleen and kidney yang deficiency by kuijiening plaster and the impacts on IFN-gamma and IL-4 contents, as well as make the comparison with oral medication of sulfasalzine (SASP). METHODS: Sixty patients of ulcerative colitis with spleen and kidney yang deficiency were randomized into a Kuijiening plaster group, a SASP group and a combined therapy group, 20 cases in each one. In the Kuijiening plaster group, Kuijiening plaster and oral administration of placebo SASP were applied. The plaster was used at Shangjuxu (ST 37), Tianshu (ST 25), Zusanli (ST 36), Mingmen (GV 4) and Guanyuan (CV 4). In the SASP group, was applied Kuijiening plaster placebo at the points and SASP oral administration was adopted. In the combined therapy group, Kuijiening plaster and SASP oral administration were given. The duration of treatment was 60 days. The follow-up visit was 2 months after treatment. The comprehensive efficacy, the efficacy on TCM syndrome and the changes in serum IFN-gamma and IL-4 before and after treatment were compared among the three groups. RESULTS: The efficacy on TCM syndrome in the Kuijiening plaster was similar to the SASP group [85.0% (17/20) vs 75.0% (15/20), P > 0.05]. The efficacy on TCM syndrome in the Kuijiening plaster group was superior to the western medicine group [80.0% (16/20) vs 60.0% (12/20), P < 0.05]. The total effective rate of the comprehensive efficacy in the combined therapy group was 95.0% (19/20) and that of TCM syndrome efficacy was 90.0% (18/20), which were all superior to the other two groups (all P < 0.05). The differences in serum IFN-gamma and IL-4 were statistically significant before and after treatment in all of the three groups (all P < 0.05). The treatments in the three groups reduced serum IFN-gamma content and increased serum IL-4 content. The results in the Kuijiening plaster group were superior to the SASP group (P < 0.05) and the results in the combined therapy group were superior to the other two groups (all P < 0.05). CONCLUSION: Kuijiening plaster is effective in the treatment of ulcerative colitis of spleen and kidney yang deficiency, which is not inferior to that of SASP. The efficacy of kuijiening plaster on relieving TCM syndrome and improving body immunity is much superior to SASP. The effect is much better with SASP combined in the treatment.

[Analysis of associated rules between Chinese medical syndrome typing features of epigastric pain and endoscopic features].

OBJECTIVE: To explore Chinese medical syndrome typing features of epigastric pain and its association with gastroscopic manifestations. METHODS: Using associated rules data mining method, an analysis study was performed in 234 patients with epigastric pain by combining holistic Chinese medical syndrome typing and results of gastroscopic examinations. RESULTS: Pi-Wei deficiency cold syndrome (PWDCS) was associated with pale gastric mucosa, or superficial ulcers accompained with unobvious redness and swelling. Inversed Gan-qi invading Wei syndrome (IGIWS) was associated with bile regurgitation. Damp-heat stagnating in the middle jiao syndrome (DSMJS) was associated with obvious redness and swollen mucosa, or ulcers accompanied with yellow and white fur. Wei-yin exhaustion syndrome (WYES) was associated with coarse mucosa and exposed blood vessels, or rare mucus. Retention of blood stasis in Wei syndrome was associated with bulged swelling and erosion. Indigestion impairing Wei syndrome (IIWS) was associated with retention of water and food, or more mucus attached. The endoscopic manifestations of PWDCS and DSMJS occurred most frequently, while PWDCS and IGIWS ranked the first place in macroscopic Chinese medical syndrome typing. CONCLUSIONS: The manifestations of endoscopic features were closely associated with Chinese medical syndrome typing. The syndrome typing features of epigastric pain could be objectively and further grasped by summarizing syndrome typing laws using modern devices such as gastroscopic examinations, thus improving the accuracy of clinical syndrome typing.

[Effects of three traditional Chinese medicine with pungent-flavor, warm-nature and meridian tropism in lung on lung-yang deficiency rats induced by compound factors].

OBJECTIVE: To observe the effects of three traditional Chinese medicine (TCM) such as Amomi Fructus Rotundus, Perillae Folium and Angelicae Dahuricae Radix on lung-yang deficiency rats induced by compound factors. METHOD: Lung-yang deficiency rats were established with three-factor combination, such as smoking (exogenous evil effect on lung), swimming in common and ice water (cold body and exhaustoin) and drinking ice water (inhale cold). Meanwhile, rats were given water extracts of the three TCM by intragastric administration for 24 days everyday. Indexes such as general behavior, weight, rectal temperature, back temperature and grip strength were observed. Blood was collected to determine NO, IgG in blood serum. Lung and heart were dissected to measure organs index. RESULT: The water extracts of Amomi Fructrs Rotundus, Perillae Folium and Angelicae Dahuricae Radix could markedly heighten weight, back temperature, grip strength, content of IgG in blood serum, reduce content of NO in blood serum, lung index and heart index. The water extracts of Amomi Fructrs Rotundus and Perillae Folium could heighten rectal temperature. CONCLUSION: Amomi Fructrs Rotundus, Perillae Folium and Angelicae Dahuricae Radix were TCM with pungent-flavor, warm-nature and meridian tropism in lung, which could improve the symptoms of physique emaciation, aversion to cold of the back, weary and acratia and so on. It provides an important reference for the regularity of the properties theories about pungent-flavor, warm-nature and meridian tropism in lung.

Long-term dietary supplementation with a yang-invigorating Chinese herbal formula increases lifespan and mitigates age-associated declines in mitochondrial antioxidant status and functional ability of various tissues in male and female C57BL/6J mice.

To investigate whether Vigconic 28 (VI-28), a Yang-invigorating Chinese herbal formula, could affect survival of aging animals, male and female C57BL/6J mice were given a VI-28-supplemented diet (0.05 and 0.5%, wt/wt) starting at 36 weeks of age, until death. VI-28 dietary supplementation at 0.05% significantly increased median lifespans of both male and female mice as compared to controls. Survival enhancement was associated with protection against age-associated impairments in mitochondrial antioxidant status and functional ability in various tissues. In conclusion, VI-28 could retard the aging process in mice, probably by mitigating age-associated declines in mitochondrial antioxidant status and functional ability in tissues.

Relationship between various Chinese medicine types and T-cell subsets in patients with ulcerative colitis.

OBJECTIVE: To investigate the relationship between various Chinese medicine (CM) types and T-cell subsets (CD4(+) and CD8(+)) in the colonic mucous membranes of patients with ulcerative colitis (UC). METHODS: Fifty UC patients were enrolled, after differentiation into four types by CM syndromes, i.e., the internal heat-damp accumulation type (IHDA), the qi-stagnancy with blood stasis type (QSBS), the Pi-Shen yang-deficiency type (PSYD) and the yin-blood deficiency type (YBD). From every patient, 3-5 pieces of intestinal mucous membrane tissues were taken through colonoscopy to determine the levels of the T-cell subsets (CD4(+) and CD8(+)) using immunohistochemical method. The results were compared with those in the normal control. RESULTS: The level of CD8(+)increased and the ratio of CD4(+)/CD8(+)decreased mainly in colonic mucous membranous tissues in UC patients. The level of CD4(+)decreased significantly in IHDA types (P<0.01), but decreased only slightly in the PSYD, QSBS and YBD types. CD8(+)increased significantly in the IHDA types (P<0.01), but only slightly in the other three types. CONCLUSION: The IHDA types of UC are closely related with T-cell subsets. The difference of T-cell subsets in various IHDA types of UC patients has provided a theoretical basis for syndrome differentiation in the CM typing of UC.

[Construction of the male rat model of adenine-induced infertility with kidney-yang deficiency].

OBJECTIVE: To investigate the time- and dose-related reproductive toxicity of adenine in male Wistar rats. METHODS: Adenine was dissolved with Arabian sol at the proportion of 1:10 and different adenine solutions were prepared at the concentration of 0, 50, 100, 150 and 200 mg/ml. Seventy-five male Wistar rats were equally assigned to 5 dose groups and received intragastric administration of the adenine solution at 1 ml/(100 g x d). Five from each group were sacrificed every 10, 20 and 30 days, their sperm quality and body and genital weight measured, pathological examinations conducted, sex hormone changes detected by radioimmunoassay, and analyses made on the time- and dose-related reproductive toxicity of adenine. RESULTS: With the increase in the time and dose of adenine administration, signs of kidney-yang deficiency gradually appeared in all the dose groups on the third day; and statistically significant changes were observed in sperm concentration and motility, and serum testosterone and luteinizing hormone levels in the 100 mg/(100 g x d) group (P < 0.05 or P < 0.01) by the 10th day. The same changes were observed in 50 mg/(100 g x d) group by the 20th day. Different degrees of pathological changes were noted in a time and dose-dependent manner in all the dose groups, suggestive of a progressive reduction of the reproductive function with the increase of time and dose of adenine administration. CONCLUSION: For the construction of the male rat model of adenine-induced infertility with kidney-yang deficiency, the best dose is 50-100 mg/(100 g x d) and the best administration time is 10-20 consecutive days intragastrically.

[Study on establishing animal model of myocardial infarction of blood-stasis and Xin-qi deficiency syndrome or blood-stasis and Xin-yang deficiency syndrome type].

OBJECTIVE: To investigate the method for establishing animal model of integrative medical disease/ syndrome and its evaluation. METHODS: Rat myocardial (heart failure) model was established by anterio-descending coronary arterial ligation, and treated by nitric oxide synthase inhibitor 9-12 weeks after operation to induce hypertension and aggravate heart failure. The model rat was observed 12 weeks to collect the information of four-diagnosis, for visceral qi-blood and excess-deficiency syndrome differentiation in combining with the eight-principal syndrome differentiation according to the standard of deficiency syndrome and blood-stasis syndrom issued by Chinese Association of integrative medicine. RESULTS: After modeling, the model rats revealed ascended ST segment and abnormal Q-wave in ECG, with the visceral ratio, left ventricular area and myocardial collagen content significantly more than those in the sham-operative group (P < 0.01), showing a blood-stasis syndrome. Besides, Xin-qi deficiency syndrome, exhibiting as quickened heart rate, shortened swimming time and lowered cardiac function, appeared in the model rats (P < 0.01), which was aggravated in the late stage due to the increased blood pressure and deteriorated cardiac function, even revealed the manifestation of Yang-deficiency syndrome as low body temperature and polyuria. CONCLUSION: The basic thinking path of TCM animal disease/syndrome model establishment and evaluation should adopt the normative, mature and unified standard and methods formed in clinical and experimental study of integrative medicine.

[Study on immunological pathogenesis of 59 patients with multiple sclerosis of different TCM syndrome types].

OBJECTIVE: To explore the immunological pathogenesis of multiple sclerosis (MS) patients of different TCM syndrome types. METHODS: Fifty-nine MS patients were assigned to two types by syndrome typing according to their clinical manifestations, the Gan-Shen yin-deficiency (GSYD, 40 cases) type and the both yin-yang deficiency (YYD, 19 cases) type. Difference of patients' age of first attack, times of relapsing, duration of disease, MRI finding and evoked potential between the two groups were compared. The immunology indexes were also compared in part of the patients (26 cases in GSYD type and 12 cases in YYD type). RESULTS: The age of first attack was later (P < 0.01), level of myelin basic protein in cerebrospinal fluid was higher (P < 0.05), in the YYD type than those in the GSYD type. Besides, the relapsing time in GSYD type, and the blood-brain barrier index and level of myelin basic protein in YYD type showed an ascending trend (P = 0.056, 0.074, 0.093, respectively). CONCLUSION: Immunological difference exists between the MS patients of GSYD type and those of YYD type.

[Study on correlation between TCM syndrome type and pathological changes of liver tissue in 260 patients with chronic hepatitis B].

OBJECTIVE: To study the correlation between TCM syndrome type and liver tissue pathological changes in patients with chronic hepatitis B (CHB) in order to provide evidence for syndrome differentiation. METHODS: Syndrome typing as well as liver pathological grading and staging of liver biopsy were performed on 260 patients with CHB, then the relationship between them was analyzed. RESULTS: (1) The grade of liver inflammation was mainly G1 and G2 in patients of Gan-qi stagnation and Pi-deficiency type (type 1); G2 in patients of inner damp-heat retention type (type II); G3 in patients of Gan-Shen yin-deficiency type (type lII) and Pi-Shen yang-deficiency type (type IV); while G4 occurred mainly in patients of blood stasis blocking collateral type (type V), showing significant difference as compared with other syndrome types. (2) The liver pathological stage in patients of type I and II was mainly S1 and S2, while S3 and S4 occurred mainly in patients of type III and type IV. (3) The pathological change was mainly G3-G4 and S3-S4 in blood stasis syndrome, while it was mainly G1-G2 and S1-S2 in non-blood stasis syndrome. CONCLUSION: The TCM syndrome type is correlated with liver tissue pathological change to certain extent, among them, syndrome with or without blood stasis showed the closest correlation. The syndrome type of CHB patients developed, along with the aggravating of liver pathological injury, from sthenia to asthenia, from qi to blood, and finally to the blood stasis blocking collateral. So the treatment should be lay stress on activating blood circulation to remove stasis, and be implemented by 3 stages.