[Acupuncture Stimulation of Acupoints of Multiple Meridians for Patients with Diminished Ovarian Reserve of Both Yin and Yang Deficiency].

OBJECTIVE: To observe the effect of acupuncture stimulation of acupoints of the Conception Vessel, Kidney Meridian, Spleen Meridian, and Bladder Meridian on menstrual cycles and duration, and serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estrogen 2 (E(2)) levels in patients with diminished ovarian reserve (DOR). METHODS: A total of 96 patients with DOR of both yin and yang deficiency were randomly divided into medication group and acupuncture group (n = 48 cases in each group). Patients of the medication group were treated by Estradiol Valerate tablets, 2 mg/d on the first 10 days, and Estradiol Cyproterone, 3 mg/d from day 11 to 21, followed by 5 -7 days' rest, and the next therapeutic course, continuously for 6 months. For patients of the acupuncture group, filiform acupuncture needles were separately inserted into every 5 points of the Conceptional Vessel, Kidney, Spleen and Bladder Meridians, manipulated with uniform reinforcing and reducing methods till Deqi, and retained for 40 min. The treatment was conducted once daily for consecutive 10 days in one menstrual cycle, beginning from the 10(th) day on after menstruation, which was repeated for 6 months. The integrative scores (normal = 0, mild=2, moderate=4 and severe=6 points) of menstrual cycle, menstrual duration, amount, color, quality [blood blot or ame- nia, symptoms of traditional Chinese medicine (TCM)] were assessed according to "Guiding Principles for Clinical Trials of New Drugs of Chinese Materia Medica". Serum FSH, LH and E(2) contents were detected by Roche's electrochemical luminescence method. RESULTS: After the treatment, of the two 48 cases in the medication and acupuncture groups, 12 (25.0%) and 20 (4.7) were cured, 11 (22.9 %) and 12 (25.0 %) experienced marked improvement in their symptoms, 20 (41.7%) and 10 (20.8%) were effective, and 5 (10.4%) and 6 (12.5%) failed, with the effective rate being 89.6% and 87.5%, respectively. The integral score of TOM symptoms, menstrual cycle, serum FSH, LH and E2 contents were considerably diminished in both groups after 6 months of treatment (P<0.05), and the TOM symptom score, menstrual cycle, and serum FSH, LH and E2 levels were significantly lower in the acupuncture group than in the medication group 6 months after cease of the treatment (P<0.05), while the menstrual duration in each cycle was notably longer in both groups after the treatment, and evidently longer in the acupuncture group than in the medication group 6 months after cease of the treatment (P<0.05). No significant differences were found between the two groups in the effective rate, score of TOM symptoms, menstrual cycle and duration, and serum FSH, LH and E(2) contents following the treatment (P>0.05). CONCLUSION: Acupuncture stimulation of acupoints of the Conception Vessel, Kidney, Spleen, and Bladder Meridians is effective in improving clinical symptoms of DOR patients with deficiency of both yin and yang, and has a longer effect, which may be closely associated with its functions in lowering serum FSH, LH and E(2) levels through regulating the hypothalamic-pituitary-ovarian axis.

Spleen-yang-deficiency patients with polycystic ovary syndrome have higher levels of visfatin.

OBJECTIVE: To study serum visfatin levels in women with polycystic ovary syndrome (PCOS) grouped by Traditional Chinese Medicine (TCM) patterns. To study the correlations of serum visfatin levels with homeostatic model assessment insulin resistance (HOMA-IR), fasting plasma glucose (FPG), fasting insulin (FINS), body mass index (BMI), testosterone (T), total cholesterol (TC), and triglycerides (TG). METHODS: Two hundred and twelve PCOS patients were placed into the following TCM pattern subgroups: Kidney-Yang deficiency (KYD) group, Spleen-Yang deficiency (SYD) group, stagnant Liver-Qi transforming into heat (SLQTH) group, and Kidney-Yin deficiency (KYIND) group. The correlations between serum visfatin levels and HOMA-IR, FPG, FINS, BMI, T, TC, and TG were analyzed. RESULTS: Of all patients with PCOS, there were 82 in the KYD group (38.6%), 67 in the SYD group (31.6%), 37 in the SLQTH group (17.5%), and 26 in the KYIND group (12.3%). Visfatin levels in all PCOS subgroups were higher than those in the control group (P < 0.01 or P < 0.05). Among these subgroups, the visfatin levels in the SYD group were significantly higher than those in the other three TCM pattern groups (P < 0.05). There were no statistical differences among the remaining three pattern groups. The levels of BMI, FINS, HOMA-IR, T, and TG were significantly higher in all subgroups than those in the control group (P < 0.05). There were no significant differences in FPG and TC between all PCOS subgroups and the control group (P > 0.05). The SYD group had higher levels of FINS and HOMA-IR compared with the KYD, SLQTH, and KYIND groups (P < 0.05). In all subgroups, after controlling for BMI, TG, TC, and age, visfatin was positively correlated with FINS (r = 0.197, P = 0.015) and HOMA-IR (r = 0.173, P = 0.033), and was not correlated with T. CONCLUSION: KYD and SYD patterns are most common in PCOS patients. Increased visfatin is a common pathophysiologic manifestation in PCOS patients. The SYD group had the highest levels of visfatin, and visfatin was positively correlated with FINS and HOMA-IR.

Metabolic profiling reveals therapeutic biomarkers of processed Aconitum carmichaeli Debx in treating hydrocortisone induced kidney-yang deficiency syndrome rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Kidney-yang deficiency syndrome (KYDS) is a diagnostic pattern in traditional Chinese medicine (TCM) and clinical data showed that the unbalance in adrenal cortical hormone is the key issue in KYDS patients. The processed Ranunculaceae aconitum carmichaeli debx (bai-fu-pian in Chinese, BFP) is one of the most commonly used Chinese herbs for treating KYDS. The present study was conducted to explore the therapeutic biomarkers of the BFP in treating hydrocortisone administration induced KYDS rats. MATERIALS AND METHODS: Thirty male Sprague-Dawley rats were randomly divided into five groups with six in each group. KYDS in rats was induced by i.p. injection of hydrocortisone at the dose of 10mg/kg per day for 15 days as described previously. The rats with KYDS were administered orally, starting from the day of hydrocortisone administration stopped, with BFP extract at the dose of 0.32g/kg, 0.64g/kg and 1.28g/kg per day respectively for 15 days. The blood samples were collected for the liquid chromatography quadruple time-of-flight mass spectrometry (LC-Q-TOF-MS) test, as well as radioimmunoassay to determine the concentrations of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and adrenocorticotrophic hormone (ACTH). The metabolic responses to BFP administration were investigated by using the principal components analysis (PCA) and orthogonal partial least squares analysis (OPLS). Bioinformatics analyses were performed by using the Ingenuity Pathway Analysis (IPA). Variance analysis and linear regression analysis were used in this study. RESULTS: The signs and concentrations of cAMP, cGMP and ACTH in the model rats were similar to those previously described about KYDS rats and BFP treatment can reverse the changes. Seventeen significantly changed metabolites among different groups were identified. Thirteen metabolites were identified in the KYDS rats comparing to healthy rats with nine up-regulated and four down-regulated. After BFP treatment at three dosages, five up-regulated metabolites including phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide were dose-dependently reversed. The network analysis with IPA showed that four canonical pathways including superpathway of methionine degradation, purine nucleotides de novo biosynthesis II, tyrosine synthesis and serotonin receptor signaling involved the therapeutic mechanism of BFP in treating the KYDS rats. CONCLUSIONS: Five therapeutic biomarkers (phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide) and two corresponding canonical pathways (amino acid metabolism and purine nucleotide metabolism) were identified to be involved in the therapeutic mechanism of BFP treating the KYDS.

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.

[Evaluation of the key indicators in the pituitary-target gland axes in the animal model with shenyang deficiency syndrome using factor analysis].

OBJECTIVE: To evaluate the key indicators in the pituitary-target gland axes in the animal model of Shen-yang deficiency syndrome (SYDS). METHODS: The 8 biological indicators [thyroid stimulating hormone (TSH), 3, 3', 5-triiodothyronine (T3), thyroxine (T4), luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), adrenocorticotropic hormone (ACTH), and cortisol (CORT)] in the pituitary-target gland axes were grouped using factor analysis. Then the sensitivity of every indicator was calculated according to the sensitivity function defined in this paper, so as to find all the most sensitive indicators in every group as key indicators of SYDS. RESULTS: The key indicators in the early period of SYDS were T, LH, T4, and CORT. The key indicators in the middle period were LH,T, CORT, and ACTH. The key indicators in the late period were LH, T, CORT, and FSH. CONCLUSIONS: T, LH, and CORT were the common key indicators of the three periods, and other different key indicator of SYDS in the early, middle and late period were T4, ACTH, and FSH respectively, which changed from the thyroid axis to the adrenal axis and then to the gonadal axis as the period changed. The key indicators in the late period were mainly in the gonadal axis, showing gonadal dysfunction in the late period.

[Regulating effect and mechanism of electroacupuncture with shuanggu yitong prescription methods on immunosenescence in aging rats with yang deficiency].

OBJECTIVE: To explore the improvement effect and the regulating mechanism of electroacupuncture with "Shuanggu Yitong" prescription methods on learning memory and immunosenescence in aging rats with yang deficiency. METHODS: Forty female Sprague-Dawley rats, 5-month-old, were randomly divided into a normal control (NC) group, an aging yang deficiency model (M) group, an electroacupuncture with Shuanggu Yitong prescription methods (EA) group and an electroacupuncture control (EAC) group, 10 rats in each group. The aging model rats with yang deficiency were established by hypodermic injection of D-galactose for 40 days and then intramuscular injection of Hydrocortisone for 7 days in the latter three groups except the NC group The EA group was treated with electroacupuncture at "Guanyuan" (CV 4), "Housanli" (ST 36) and "Baihui" (GV 20), and the EAC group was treated with electroacupuncture at "Zhongji" (CV 3), "Yinlingquan" (SP 9) and "Yintang" (EX-HN 3), six times per week for 4 weeks. The escape latency was examined by Morris water maze from the first day of the 4th week. After completion of electroacupuncture treatment, the rate of spleen lymphocyte apoptosis and the serum content of tumor necrosis factor-a (TNF-alpha) were examined by flow cytometry and enzyme linked immunosorbent assay (ELISA) technique respectively, the differences among all the groups above were compared. RESULTS: In comparison with the NC group, the escape latency [(25.4 +/- 3. 6)s vs. (16.23 +/- 2.3)s], the rate of spleen lymphocyte apoptosis [(27.25 +/- 3.3)% vs. (13.2 +/- 3.1)%] and the serum content of TNF-alpha [(15.54 +/- 3.56) pg/mL vs (7.35 +/- 2.89) pg/mL] in the M group those were all significantly increased (all P < 0.01). In the EA group, the escape latency of (17.42 +/- 3.9)s, the rate of spleen lymphocyte apoptosis of (17.2 +/- 3.25)% and the serum content of TNF-alpha of (9.51 +/- 3.53) pg/mL were all significantly lower than those in the M group (all P < 0.01) and the EAC group (all P < 0.05). CONCLUSION: Electroacupuncture with "Shuanggu Yitong" prescription can significantly improve the spatial learning and memory. The protective mechanism is related with the action of electroacupuncture in lowering the rate of spleen lymphocyte apoptosis and the serum content of TNF-alpha, and the therapeutic effect of electroacupuncture at "Guanyuan" (CV 4), "Housanli" (ST 36) and "Baihui" (GV 20) is superior to that of electroacupuncture at "Zhongji" (CV 3), "Yinlingquan" (SP 9) and "Yintang" (EX-HN 3).

[Effects of Wenshen Huatan Recipe on the renal function and foe blood lipids of patients with early-middle stage chronic renal insufficiency of Pi-Shen yang deficiency].

OBJECTIVE: To study the clinical effectiveness of Wenshen Huatan Recipe (WHR) in treating early-middle stage chronic renal insufficiency (CRI) of Pi-Shen yang deficiency syndrome (PSYDS). METHODS: Ninety early-middle stage CRI patients of PSYDS were recruited and randomly assigned to the treated group (60 cases) and the control group (30 cases) in the ratio of 2:1. All patients received conventional and symptomatic treatment, but those in the treated group received WHR additionally. The observation was lasted for six months. The total efficacy, the Chinese medicine syndrome integral, and indices of the renal functions [including serum creatinine (SCr), blood urea nitrogen (BUN), creatinine clearance (CCr), serum cystatin C (Cys-C)], blood lipids [total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C)], as well as adverse reactions were observed before and after treatment. RESULTS: After six months of treatment, the total effective rate in treated group was 70.0% (42/60), which was obviously higher than that in the control group [43.3% (13/30), P < 0.05]. The Chinese medicine symptoms in the treated group were obviously improved, showing statistical difference between the two groups (P < 0.01). Better effects in lowering SCr, BUN, Cys-C, and LDL-C and increasing CCr were obtained in the treated group than in the control group (P < 0.05). CONCLUSION: WHR could postpone the progression of CRI, improve the lipid metabolism, indicating certain therapeutic efficacy could be obtained in treating early-middle stage CRI from sputum theory.

[Effect of epimedii folium processed with different refining temperatures and amounts of sheep's oil on kidney-yang deficiency rats].

OBJECTIVE: To investigate the effects of refining temperatures and amounts of sheep's oil used in processing Epimedii Folium on Kedney-yang deficiency rats. METHOD: The corticosterone was subcutaneous injected to establish the kidney yang deficiency rat model. With the temperatures and amounts of sheep's oil were 250 degrees C/30%, 120 degrees C/30% and 120 degrees C/20% respectively, the crude drug and three kinds of pressed Epimedii Folium were extracted by water and used as examined samples while total flavonoid of Epimedii Folium was used as positive control. After examined samples and control samples were intragastirc administrated, the pharmacologic action was analyzed. RESULT: As compared to crude drug, all of the aqueous extracts of processed Epimedii Folium have stronger effect of warming kidney and enhancing yang, especially the Epimedii Folium processed by sheep's oil with refining temperatures 120 degrees C and amounts of sheep's oil 30%. Its mechanism might be related to improving the insufficiency of hypothalamic-pituitary-adrenal-thymus (HPAT) axis suppression. CONCLUSION: The refining temperature of sheep's oil can affect the quality of excipients and processed drugs. The results may be useful in explaining the mechanism of Epimedii Folium processing and establishment of pharmaceutical standard of sheep's oil used as processing excipients.

[Clinical observation on acupoint irradiation with combined laser or red light on patients with knee osteoarthritis of yang deficiency and cold coagulation type].

OBJECTIVE: To explore the effects of combined laser on patients with knee osteoarthritis (OA) of yang deficiency and cold coagulation type. METHODS: Forty-one cases with knee OA of yang deficiency and cold coagulation type were randomly divided into a combined laser group (n = 22) and a red light group (n = 19), with combined laser and red light irradiation on Dubi (ST 35) and Neixiyan (EX-LE 4), respectively. They were treated for 6 weeks. The scores of Western Ontario and McMaster Universities' Osteoarthritis Index (WOMAC VA3. 1) were used for assessment of therapeutic effects. RESULTS: After treatment for 2 weeks and 6 weeks, the WOMAC scores significantly decreased in the two groups as compared with those before treatment (P < 0.05, P < 0.01 and P < 0.001). The mean improvement rate of WOMAC scores in the combined laser group was better than those in the red light group. CONCLUSION: Combined laser irradiation can improve the pain, stiffness and functional limitation of patients with knee OA of yang deficiency and cold coagulation type.

[Diuretic effect of liniaoling(KLP)].

Experimental results have shown that KLP taken orally helps promote the diuretic action of normal Wistar rats, decrease the urine volume of debilitated rats and increase the urine volume of BPH rats. It has also been shown that KLP(i.v.) can increase the urine volume of anesthetic rats and reduce the remnant urine. In addition, KLP can contract the smooth muscle of rats and rabbits.

[Effect of zhuchun pill on immunity and endocrine function of elderly with kidney-yang deficiency].

After randomly dividing 45 elderly males with Kidney-Yang Deficiency into Zhuchun Pill (ZCP) group and placebo (starch capsule) group by single blind method, and treating them for 3 months, we found that ZCP could greatly improve the main symptoms of Kidney-Yang Deficiency, P < 0.01 by comparison with control group: the 3H-TdR lymphocyte transformation (3H-TdR LCT) rate, level of serum C3, IgA, IgD, IgG & IgM went up (P < 0.01); contents of plasma adrenocortical hormone (ACTH), testosterone (T) and cyclic adenosine monophosphate (cAMP) increased (P < 0.01-0.001): the level of superoxide dismutase (SOD) in serum also increased while lipid peroxide (LPO) in plasma decreased (P < 0.01). But for control group, there were no remarkable changes after treatment, so it revealed that the improvement of Kidney-Yang Deficiency by ZCP is realized through regulating the organism's immunity and endocrine function as well as through scavenging the free radical agents.

[Influence of stimulating zusanli with moxibustion of different quality and quantity on gastrointestinal motor function of reserpinized rats].

This article is focused on the observation of changes in body temperature, body weight, cholinesterase activity in blood, and gastrointestinal motility of reserpinized rats treated by stimulating Zusanli (ST 36) with moxibustion of different quality. (mugwort floss or pipe tobacco) and quantity (strong stimulation or weak stimulation). The experiment shows that better results are achieved with moxibustion not by burning tobacco; the result of strong stimulation with moxa-sticks is better than that of weak stimulation with the same material. Strong stimulation with moxa-sticks can obviously increase the activity of cholinesterase (P < 0.05), inhibit hyperactive gastrointestinal motility (P < 0.05), maintain normal body temperature (P < 0.05), and prevent body weight lossing. All these show that therapeutic results of moxibustion are closely related to the quality and quantity of moxibustion.