[Influence of acupuncture combined with rehabilitation therapy on limb motor function and serum cAMP and cGMP in patients with hemiplegia of cerebral ischemic stroke].

OBJECTIVE: To evaluate the therapeutic effect of the combined treatment with balance acupuncture therapy and exercise re-learning rehabilitation therapy and the impact on serum cAMP and cGMP in the patients with hemiplegia of cerebral ischemic stroke. METHODS: A total of 90 patients of hemiplegia of cerebral ischemic stroke were randomized into an observation group and a control group, 45 cases in each one. All of the patients in the two groups received health education, diet guidance, routine symptomatic treatment as well as exercise re-learning rehabilitation therapy. Additionally, in the observation group, balance acupuncture therapy was applied, in which, the acupoints on the yang aspect of the human body, on the governor vessel and bladder meridian were adopted in the morning and those on the yin aspect of the human body, on the conception vessel and kidney meridian were stimulated in the afternoon. In the control group, the regular acupuncture was given. In the two groups, both acupuncture and rehabilitation therapies were given 5 days a week, 2 week-treatment as one course and totally 2 courses were required. Separately, before and after treatment, the score of Fugl-Meyer assessment (FMA) and the score of Chinese stroke scale (CSS) were recorded, the levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) detected in serum and the clinical therapeutic effect were evaluated in the two groups. RESULTS: After treatment, FMA score was increased in the patients of either of the groups as compared with that before treatment (P<0.01) and CSS score decreased as compared with that before treatment (P<0.01). After treatment, FMA score in the observation group was higher than that in the control group (P<0.01) and CSS score was lower than the control group (P<0.01). After treatment, the level of serum cAMP of the patients in either of the groups was increased as compared with that before treatment (P<0.01) and that of cGMP decreased as compared with that before treatment (P<0.01). After treatment, the level of cAMP in the observation group was higher than that in the control group (P<0.01) and that of cGMP was lower than the control group (P<0.01). The total effective rate was 93.3% (42/45) in the observation group, better than 73.3% (33/45) in the control group (P<0.01). CONCLUSION: The balance acupuncture therapy combined with exercise re-learning rehabilitation effectively improves the motor function of the affected limb, relieves injury and regulate the levels of serum cAMP and cGMP in the patients with hemiplegia of ischemic stroke.

Analgesic action of Rubimaillin in vitro and in vivo.

Pain, a common symptom in clinics, is a serious impediment to quality of life. The analgesic drugs presently in use have poor efficacy, and are associated with undesirable side effects. Rubimaillin (Rub) is a naphthoquinone compound extracted from Chinese herbal medicine, and it has various biological activities. In this study, the analgesic effect of Rub, and its mechanism of action were investigated using glacial acetic acid-induced mice writhing model and a mice model of neurogenic and inflammatory bipolar pain. Analgesic effects were measured in different experimental groups. In vitro, RAW 264.7 cells were used to investigate the release of nitric oxide (NO), iNOS and COX-2 protein in RAW 264.7 cells stimulated with lipopolysaccharide (LPS). The results revealed that Rub reduced the number of acetic acid-induced writhing in mice, inhibited formalin-induced biphasic pain response, and suppressed the production of NO in RAW 264.7 cells. The mechanisms involved in the analgesic and anti-inflammatory effects of rub may be related to the inhibition of cyclooxygenase-2 (COX-2), endogenous inflammatory mediators, and reduction in the content of pain-induced mediators.

Total flavonoid extract of Epimedium herb increases the peak bone mass of young rats involving enhanced activation of the AC10/cAMP/PKA/CREB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium sagittatum brevicornum Maxim. is an important traditional Chinese herb that has long been used to promote bone fracture healing and treat osteoporosis. AIM OF THE STUDY: Achieving peak bone mass by adolescence has now been accepted to be fundamental for preventing osteoporosis in adulthood life. This study investigated the possibility of increasing peak bone mass in young rats using the total flavonoid extract of Epimedium herb (TFE). MATERIALS AND METHODS: TFE was intragastrically administered to one-month-old Wistar rats at a low (100 mg/kg), middle (200 mg/kg) or high dose (400 mg/kg). Whole body bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry every two weeks. When BMD of any one of TFE groups was found to be significantly higher than that of the control, all rats were sacrificed, serum samples were collected for bone turnover biochemical assays, and femurs, tibiae and vertebrae were isolated and used in BMD, mechanical, micro-structural, histomorphometric and mechanistic studies. RESULTS: Administration of TFE at middle and high doses for two months significantly increased the whole body, femoral and vertebral BMDs, and improved the bone mechanical and micro-architectural properties. The serum turnover biochemical results and the enhanced expression levels of bone-formation regulatory genes (Runx-2, OSX, and BMP-2) demonstrated that TFE administration increased bone formation but had no effect on bone resorption. The increased phosphorylation levels in femurs of PKA and CREB and expression of AC10 (the only soluble form of adenylyl cyclase) and the increased serum cAMP level after 4 h of TFE administration indicated that TFE promoted bone formation by activating the AC10/cAMP/PKA/CREB pathway in vivo. CONCLUSIONS: Oral administration of TFE at 200 mg/kg for two months can increase the peak bone mass of growing rats, suggesting the possibility of using total flavonoid extract of Epimedium herb to increase the peak bone mass in adolescence which is important for preventing osteoporosis in adult life.

12(S)-HETrE, a 12-Lipoxygenase Oxylipin of Dihomo-γ-Linolenic Acid, Inhibits Thrombosis via Gαs Signaling in Platelets.

OBJECTIVE: Dietary supplementation with polyunsaturated fatty acids has been widely used for primary and secondary prevention of cardiovascular disease in individuals at risk; however, the cardioprotective benefits of polyunsaturated fatty acids remain controversial because of lack of mechanistic and in vivo evidence. We present direct evidence that an omega-6 polyunsaturated fatty acid, dihomo-γ-linolenic acid (DGLA), exhibits in vivo cardioprotection through 12-lipoxygenase (12-LOX) oxidation of DGLA to its reduced oxidized lipid form, 12(S)-hydroxy-8Z,10E,14Z-eicosatrienoic acid (12(S)-HETrE), inhibiting platelet activation and thrombosis. APPROACH AND RESULTS: DGLA inhibited ex vivo platelet aggregation and Rap1 activation in wild-type mice, but not in mice lacking 12-LOX expression (12-LOX(-/-)). Similarly, wild-type mice treated with DGLA were able to reduce thrombus growth (platelet and fibrin accumulation) after laser-induced injury of the arteriole of the cremaster muscle, but not 12-LOX(-/-) mice, supporting a 12-LOX requirement for mediating the inhibitory effects of DGLA on platelet-mediated thrombus formation. Platelet activation and thrombus formation were also suppressed when directly treated with 12(S)-HETrE. Importantly, 2 hemostatic models, tail bleeding and arteriole rupture of the cremaster muscle, showed no alteration in hemostasis after 12(S)-HETrE treatment. Finally, the mechanism for 12(S)-HETrE protection was shown to be mediated via a Gαs-linked G-protein-coupled receptor pathway in human platelets. CONCLUSIONS: This study provides the direct evidence that an omega-6 polyunsaturated fatty acid, DGLA, inhibits injury-induced thrombosis through its 12-LOX oxylipin, 12(S)-HETrE, which strongly supports the potential cardioprotective benefits of DGLA supplementation through its regulation of platelet function. Furthermore, this is the first evidence of a 12-LOX oxylipin regulating platelet function in a Gs α subunit-linked G-protein-coupled receptor-dependent manner.

Opposing needling promotes behavior recovery and exerts neuroprotection via the cAMP/PKA/CREB signal transduction pathway in transient MCAO rats.

The aim of the present study was to investigate whether the cyclic adenosine 3',5'­monophosphate (cAMP)/protein kinase A(PKA)/cAMP­responsive element binding protein (CREB) signal transduction pathway triggered by γ­aminobutyric acid class B (GABA(B)) receptor activation is involved in neuroprotection against ischemia and behavioral recovery induced by opposing needling (ON). A total of 80 rats were randomly divided into four groups: A sham operation group, an ischemia group, an ON group and an ON group effectively inhibited by the GABA(B) receptor antagonist, CGP35384 (n=20/group). The behavior of the rats was assessed by their neurological deficit score, whereas the impairment of gait was examined using the CatWalk system. The volume of cerebral infarction was examined upon treatment with 2,3,5­triphenyltetrazolium chloride. The expression levels of CREB, GABA(B1) and GABA(B2) were examined by western blotting and reverse transcription­quantitative polymerase chain reaction, and the activity of adenylyl cyclase (AC), cAMP and PKA in the serum was detected using an enzyme­linked immunosorbent assay. In the present study, in comparison with other groups, the ON group exhibited a reduced score for the neurological deficit, the stride length and swing speed were improved, and the volume of infarction was reduced. However, these effects were reversed upon administration of CGP35384. Additionally, the expression levels of CREB, GABA(B1) and GABA(B2) were increased in the ON group. The levels of AC, cAMP and PKA in the serum were also increased in the ON group, whereas the addition of CGP35384 reversed these effects. The results of the present study demonstrated that ON markedly protected the brain against transient cerebral ischemic injury, and this effect was possibly mediated by the activation of the GABAB/cAMP/PKA/CREB signal transduction pathway. These findings implied that ON may be a potential therapeutic method for treating stroke.

Tetramethylpyrazine identified by a network pharmacology approach ameliorates methotrexate-induced oxidative organ injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Tetramethylpyrazine (TMP) is one of the active constituents extracted from a frequently used herb, Ligusticum wallichii Franchat (Chuan-Xiong in Chinese), in traditional Chinese medicine. TMP can exert multiple pharmacological actions such as anti-inflammatory, anti-oxidative damage, anti-platelet and neuroprotective effects, and its applications deserve further explored. AIM OF THE STUDY: This study aimed to determine the new role of TMP identified by a network pharmacology approach to alleviate the methotrexate (MTX)-induced oxidative injury and characterize their mechanism of combinational actions. MATERIALS AND METHODS: A network pharmacology-based screening strategy is applied for target profile prediction and pharmacological characterization of herbal compounds, which is used to guide the following in vitro and in vivo experiments. The effect of herbal compounds identified by network pharmacology approaches to reduce the toxicity of MTX was assessed by MTX-induced rat toxicity model. The potential targets of TMP in this study were evaluated using standard protocols provided by Cerep, Inc. RESULTS: This strategy identified TMP from Ligusticum wallichii Franchat as a potent compound for ameliorating the oxidative organ injury of MTX. According to the predicted target profiles of TMP, a possible mechanism of the abrogation of MTX-induced toxicity is that TMP could upregulate cAMP by inhibiting phosphodiesterase (PDE) 10A2 activity. Another novel finding is that the competitive binding and antagonistic effects of TMP on adenosine receptor 2A and 2B appear to play important roles in the TMP-mediated reversal of MTX-induced hepatic injury. CONCLUSION: TMP identified by a network pharmacology approach could ameliorate MTX-induced oxidative organ injury. This study provides important evidence for the preclinical evaluation of TMP and MTX as a novel combinatorial remedy.

Modulation of 3',5'-cyclic AMP homeostasis in human platelets by coffee and individual coffee constituents.

3',5'-Cyclic AMP (cAMP) is one of the most important second messengers in mammalian cells, mediating a multitude of diverse cellular signalling responses. Its homeostasis is primarily regulated by adenylate cyclases and phosphodiesterases (PDE), the activities of which are partially dependent on the downstream events of adenosine receptor signalling. The present study was conducted to determine whether coffee constituents other than caffeine can influence the homeostasis of intracellular cAMP in vitro and in vivo by evaluating the effects of selected constituents present in coffee, coffee brews and coffee extracts on platelet PDE activity. In addition, to evaluate the potential effects of these constituents on platelet cAMP concentrations and PDE activity in humans, a 7-week pilot intervention study with eight subjects was conducted. The subjects consumed a regular commercial coffee and a low-caffeine coffee at a rate of 750 ml/d for 2 weeks each. The in vivo results revealed a highly significant inhibition of PDE activity (P< 0·001) after coffee intervention that was not directly dependent on the caffeine content of coffee. Although our in vitro and in vivo findings suggest that caffeine plays some role in the modulation of platelet cAMP status, other natural and roasting-associated compounds such as pyrazines and other currently unidentified species also appear to contribute significantly. In conclusion, moderate consumption of coffee can modulate platelet PDE activity and cAMP concentrations in humans, which may contribute to the putative beneficial health effects of coffee. Further detailed mechanistic investigations will be required to substantiate these beneficial effects and to elucidate the underlying mechanisms.

[Study on the regulation role of semen persicae to cAMP-PKA signal pathway in the rats with cold and heat blood stasis syndrome].

OBJECTIVE: To study the regulation role of with neutral property to cAMP-PKA pathway in the rats with cold and heat blood stasis syndromes and it's mechanism. METHODS: 60 rats were randomly divided into normal control group, model control group, Semen Persicae group, radix salvia miltiorrhiza group, rhizoma chuanxiong group, 12 rats per group. The three herb groups were orally given relative herbs decoction, whose dosages were equal to 10 times the human clinical dose, normal and model control groups were orally given water, 2 times/day, 20 mL/kg, for 7 days. Experiments in rats with cold and heat blood stasis syndromes were carried on respectiverly. In heat blood stasis syndromes, except normal control group, the other groups were intraperitoneally injected 10% carrageenan, 5 mL/kg, 1 times/day, for 3 days;24 hours after the last injection, subcutaneously injected 20% dry yeast suspension, 10 mL/kg. In cold blood stasis syndromes, except normal control group, the other groups were put into fridge, temperature--(18 +/- 2) degrees C, 2 hours/ times, 2 times/day, for 7 days. Separately draw 5 ml abdominal aortic blood and taken abdominal aorta, 6 hours and 12 hours after finishing the model in the two syndromes. Tested the cAMP content by elisa, tested the PKA protein expression by Western blot. RESULTS: Semen Persicae with neutral property, could decrease the content of cAMP in plasma (P < 0.01), inhibit the expression of protein PKA (P < 0.05) in rats with heat stagnation and blood stasis syndrome, increase the plasma content of cAMP (P < 0.01) and increase the expression of protein PKA (P < 0.01) in rats with cold stagnation and blood stasis syndrome. Semen Persicae had two-way adjustment action on CAMP-PKA signal pathway. CONCLUSION: In different internal environment of heat stagnation and blood stasis syndrome, cold stagnation and blood stasis syndrome, Semen Persicae with neutral property has two-way adjustment to cAMP-PKA signaling channel, which may be one of the mechanism of it's two-way application.

In African American type 2 diabetic patients, is vitamin D deficiency associated with lower blood levels of hydrogen sulfide and cyclic adenosine monophosphate, and elevated oxidative stress?

African Americans (AA) have a higher incidence of cardiovascular disease and vitamin D (VD) deficiency compared with Caucasians. Hydrogen sulfide (H(2)S) is an important signaling molecule. This study examined the hypothesis that blood levels of H(2)S are lower in AA type 2 diabetic patients (T2D). Fasting blood was obtained from T2D and healthy controls. Results showed a significant decrease in plasma levels of cyclic adenosine monophosphate (cAMP) and H(2)S in AA T2D but not in Caucasian T2D when compared with those of respective age- and race-matched healthy controls. Plasma VD levels were significantly lower in AA T2D compared with Caucasian T2D. Cell culture studies demonstrate that 1,25(OH)(2)-VD supplementation significantly increased expression of cystathionine-γ-lyase (CSE), H(2)S formation, and cAMP secretion, but decreased reactive oxygen species in high glucose-treated U937 monocytes. This suggests that VD supplementation upregulates CSE and H(2)S formation and decreases oxidative stress, and that VD deficiency may contribute to the malfunctioning of H(2)S signaling and thus a higher incidence of vascular inflammation in AA. These results lead to the hypothesis that VD supplementation can replenish blood concentrations of H(2)S and cAMP and lower oxidative stress and cardiovascular disease in AA T2D.

The inhibitory in-vitro effect of high-dose levosimendan on platelet function may be mediated through its action as a phosphodiesterase inhibitor.

OBJECTIVE: Levosimendan enhances cardiac contractility by increasing myocyte sensitivity to calcium and causing vasodilatation. Although studies have evaluated the efficacy of levosimendan in heart failure, whether levosimendan produces an effect on platelets is a subject of controversy. In the present study, the in-vitro effect of levosimendan on platelet aggregation was investigated. The effect of levosimendan on the cyclic AMP concentration was determined according to its second mode of action as a selective phosphodiesterase III inhibitor. MATERIALS AND METHODS: Platelet aggregation setting was performed using venous blood from three healthy volunteers. Different concentrations of levosimendan solution were prepared that would result in 0.04-125 µg/ml levosimendan concentrations in whole blood and in platelet-enriched plasma. After incubation for 3 min at 37°C, aggregation responses were evaluated with ADP (10 µmol/l), collagen (5 µg/ml), or NaCl. The cyclic AMP concentration was determined using the enzyme-linked immunosorbent assay technique. RESULTS: The in-vitro results clearly showed that there was only a relationship between a high levosimendan concentration (12-125 µg/ml) and inhibition of platelet aggregation that was negatively dependent on the cAMP concentration. CONCLUSION: Levosimendan has no significant effect as a phosphodiesterase III inhibitor on in-vitro platelet aggregation in clinically relevant doses.

Haemodynamic, endocrine and renal actions of adrenomedullin 5 in an ovine model of heart failure.

AM5 (adrenomedullin 5), a newly described member of the CGRP (calcitonin gene-related peptide) family, is reported to play a role in normal cardiovascular physiology. The effects of AM5 in HF (heart failure), however, have not been investigated. In the present study, we intravenously infused two incremental doses of AM5 (10 and 100 ng/min per kg of body weight each for 90 min) into eight sheep with pacing-induced HF. Compared with time-matched vehicle control infusions, AM5 produced progressive and dose-dependent increases in left ventricular dP/dt(max) [LD (low dose), +56 mmHg/s and HD (high dose), +152 mmHg/s] and cardiac output (+0.83 l/min and +1.81 l/min), together with decrements in calculated total peripheral resistance (-9.4 mmHg/min per litre and -14.7 mmHg/min per litre), mean arterial pressure (-2.8 mmHg and -8.4 mmHg) and LAP (left atrial pressure; -2.6 mmHg and -5.6 mmHg) (all P<0.001). HD AM5 significantly raised PRA (plasma renin activity) (3.5-fold increment, P<0.001), whereas plasma aldosterone levels were unchanged over the intra-infusion period and actually fell in the post-infusion period (70% decrement, P<0.01), resulting in a marked decrease in the aldosterone/PRA ratio (P<0.01). Despite falls in LAP, plasma atrial natriuretic peptide and B-type natriuretic peptide concentrations were maintained relative to controls. AM5 infusion also induced significant increases in urine volume (HD 2-fold increment, P<0.05) and urine sodium (2.7-fold increment, P<0.01), potassium (1.7-fold increment, P<0.05) and creatinine (1.4-fold increment, P<0.05) excretion and creatinine clearance (60% increment, P<0.05). In conclusion, AM5 has significant haemodynamic, endocrine and renal actions in experimental HF likely to be protective and compensatory in this setting. These results suggest that AM5 may have potential as a therapeutic agent in human HF.

Ent-16ß,17-dihydroxy-kauran-19-oic acid, a kaurane diterpene acid from Siegesbeckia pubescens, presents antiplatelet and antithrombotic effects in rats.

The antiplatelet and antithrombotic effects of ent-16ß,17-dihydroxy-kauran-19-oic acid (DDKA) isolated from Siegesbeckia pubescens were investigated with different methods both in vitro and in vivo. We tested the antithrombotic activity of DDKA in arterio-venous shunt model. The effects of DDKA on adenosine diphosphate (ADP)-, Thrombin-, Arachidonic acid-induced rat platelets aggregation were tested in vitro. We also assessed its bleeding side effect by measuring coagulation parameters after intravenous administration for 5 days and investigated the potential mechanisms underlying such activities. In vivo, DDKA significantly reduced thrombus weight in the model of arterio-venous shunt. Meanwhile, DDKA increased plasma cAMP level determined by radioimmunoassay in the same model. Notably, DDKA prolonged PT and APTT in rats after intravenous administration DDKA for successive 5 days. In vitro, pretreatment with DDKA on washed rat platelets significantly inhibited various agonists stimulated platelet aggregation and caused an increase in cAMP level in platelets activated by ADP. These findings support our hypothesis that DDKA possesses antiplatelet and antithrombotic activities. The mechanisms underlying such activities may involve the anticoagulatory effect and cAMP induction.

[Effect and mechanism of reserpine for changing salivary protein secretion in Pi-deficient rats].

OBJECTIVE: To study the effect of reserpine (RSP) for changing salivary protein secretion in Pi-deficient rats and to explore its possible mechanism. METHODS: Twenty rats allocated in the RSP group were given subcutaneous injection of RSP [0.4 mg/(kg x d)] for 9 successive days, while the other 20 rats in the control group were injected with same volume of saline instead. On the 10th day, ten rats randomly selected from each group were subjected for extracting saliva to detect salivary amylase activity (sAA) before and after an acid stimulation; and drawing blood from the orbital vein to measure the contents of vasoactive intestinal peptide (VIP) and cyclic adenosine monophosphate (cAMP). Then they were sacrificed and their parotids were taken out for pathological examination with HE staining, as well as for VIP and cAMP measuring, and zymogen granules counting under a transmission electron microscope. The remainder animals were stopped injecting and normally fed to 40 days, then subjected to be detected as above-mentioned. RESULTS: Food intake and body weight reduction were more significantly in the RSP group than in the control group. On the 10th day, the ratio of sAA before/after stimulation in the RSP group was 0.39 +/- 0.18, significantly lower than that in the control group (0.80 +/- 0.21, P < 0.01), but it was restored rapidly, reaching the normal range on the 25th day, on the 40th day, it became significantly different to the level on the 10th day (P < 0.05) and approached the level in the control group (P > 0.05). No significant pathological change of parotid was found in both groups; but the number of zymogen granules in the RSP group was remarkably more than that in the control group (41.4 +/- 4.9 vs 34.6 +/- 5.2, P < 0.01). Serum level of VIP in the RSP group was significantly less while that of cAMP was higher than that in the control group (22.5 +/- 13.1 mg/L vs 38.5 +/- 14.1 mg/L, and 125.8 +/- 15.5 micromol/L vs 105.3 +/- 16.7 micromol/L, both P < 0.05), but no inter-group difference was found in parotid tissue contents of both VIP and cAMP. All the indices detected became equivalent in the two groups on the 40th day. CONCLUSION: The reduction of salivary protein in Pi-deficient rats induced by RSP may be related to the regulatory pathway of VIP and cAMP.

Effects of polydatin on attenuating ventricular remodeling in isoproterenol-induced mouse and pressure-overload rat models.

The study was designed to examine the effects of polydatin on ventricular remodeling induced by isoproterenol in mice and by abdominal aortic banding in rats. Polydatin reduced cardiac weight indexes in mice and rats, lowered the contents of cyclic AMP and angiotensin II in mice. It also decreased the size of cardiomyocyte, the levels of aldosterone, tumor necrosis factor-α, angiotensin II and endothelin-1, reduced ventricular collagen volume and decreased blood pressure in rats. The results demonstrate that polydatin has the beneficial effects on attenuating ventricular remodeling, which are associated with its inhibiting the activation of neurohormone, especially in rennin-angiotensin-aldosterone system.

Lower-extremity functional electrical stimulation decreases platelet aggregation and blood coagulation in persons with chronic spinal cord injury: a pilot study.

BACKGROUND: Individuals with spinal cord injury (SCI) develop premature cardiovascular disease. Regular exercise reduces the incidence and symptoms of cardiovascular disease in able-bodied individuals; these salutary effects of exercise have not been documented in persons with SCI. OBJECTIVE: To evaluate the effects of functional electrical stimulation leg cycle ergometry (FES-LCE) exercise training on platelet aggregation and blood coagulation in persons with SCI. PARTICIPANTS: Subjects (n=14) with stable chronic (>1 year) paraplegia (T1-T10) or tetraplegia (C4-C8). METHODS: Blood samples were collected before and after the first and eighth sessions (2 sessions per week for 4 weeks) of FES exercise. RESULTS: Platelet aggregation was inhibited by 20% after the first session and by 40% (P < 0.001) after the eighth session. Thrombin activity was unchanged after the first session (10.7 +/- 0.85 s to 10.43 +/- 0.56 s) and decreased after the eighth session (12.5 +/- 1.98 s to 11.1 +/- 1.7 s; P < 0.0003). Antithrombin III activity increased after the first (103.8% +/- 8.9% to 110% +/- 6.9%; P < 0.0008) and eighth sessions (107.8% +/- 12.1% to 120.4% +/- 13.1%; P < 0.0001). Cyclic adenosine monophosphate increased after the first (9.9% + 2.5% to 15.8% +/- 3%; P < 0.001) and eighth sessions (17.8% +/- 4.2% to 36.5% +/- 7.6%; P < 0.0001). After the eighth session, factors V and X increased significantly (88% +/- 27% to 103% +/- 23%, P < 0.0001; 100% +/- 40% to 105% +/- 7%, P < 0.01, respectively); factors VII and VIII and fibrinogen did not change significantly. A significant reduction in platelet activation/aggregation was demonstrated in response to FES-LCE. The decrease in thrombin level was caused by the simultaneous increase in antithrombin activity. CONCLUSION: These findings provide new insight into the potential protective effects of FES-LCE against the risk of cardiovascular disease.

[Effects of Yiniao Recipe on serum antidiuretic hormone level and plasma ratio of cAMP to cGMP in rats with kidney-yang deficiency].

OBJECTIVE: To investigate the effects of Yiniao Recipe, a compound traditional Chinese herbal medicine, on contents of serum antidiuretic hormone, and plasma cyclic adenosine monophosphate and cyclic guanosine monophosphate in rats with kidney-yang deficiency. METHODS: Forty male Wistar rats were randomly divided into blank control group, untreated group, desmopressin (Minirin) group, low-dose Yiniao Recipe group and high-dose Yiniao Recipe group, with 8 rats in each group. Rats in the blank control group were injected with 0.2 mL normal saline, and rats in the other groups were given intramuscular injection of hydrocortisone 25 mg/kg, 1 time daily for 21 consecutive days; from the 8th day of injection, rats were given double distilled water, Minirin, and high- and low-dose Yiniao Recipe respectively for 30 days. Before and after treatment, 24-hour urine volume was observed, and serum antidiuretic hormone (AVP) as well as plasma cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) contents were detected by enzyme-linked immunosorbent assay. The cAMP/cGMP ratio and morphological changes in renal tissues were also observed. RESULTS: Compared with blank control group, 24-hour urine volume, serum AVP content and cAMP/cGMP ratio in the untreated group were decreased; compared with the untreated group, Minirin and Yiniao Recipe at low and high doses reduced 24-hour urine volume and increased serum AVP content and cAMP/cGMP ratio significantly (P<0.05 or P<0.01). There were no obvious pathological changes in renal tissue in all groups. CONCLUSION: Yiniao Recipe may reduce 24-hour urine volume by increasing serum AVP content and regulating the ratio of cAMP to cGMP in kidney-yang deficiency rats.

Effect of sodium houttuyfonate on myocardial hypertrophy in mice and rats.

OBJECTIVES: The aim of the study was to determine the effect of sodium houttuyfonate on myocardial hypertrophy and its mechanism of action in mice and rats. METHODS: A mouse model of myocardial hypertrophy was established by subcutaneous injection with isoproterenol. Mice were randomly divided into five groups: normal control; isoproterenol control; isoproterenol plus metoprolol; isoproterenol plus low- and high-dose sodium houttuyfonate. A rat model of myocardial hypertrophy was established by intraperitoneal injection with L-thyroxine. Rats were randomly divided into five groups: normal control; L-thyroxine control; L-thyroxine plus captopril; L-thyroxine plus low- and high-dose sodium houttuyfonate. At the end of the experiments, the left ventricular weight index and heart weight index were determined in mice and rats, the size of cardiomyocytes was measured in rats and the concentrations of cAMP in plasma and angiotensin II in ventricular tissue of mice were detected by radioimmunoassay. The endothelin-1 concentration was measured by radioimmunoassay and the hydroxyproline content was measured by a digestive method in ventricular tissue of rats. KEY FINDINGS: After 7-9 days of treatment, sodium houttuyfonate significantly reduced the left ventricular weight index and heart weight index in mice and rats with myocardial hypertrophy, decreased the size of cardiomyocytes in rats, and reduced the content of cAMP and angiotensin II in mice with myocardial hypertrophy. It also decreased the endothelin-1 concentration and the hydroxyproline content in ventricular tissue in rats. CONCLUSIONS: Sodium houttuyfonate can inhibit myocardial hypertrophy in mouse and rat models by restricting the activity of the sympathetic nervous system and decreasing the levels of angiotensin II and endothelin-1 in ventricular tissue.

[Changes of endocrine and immune function in subjects of yang deficiency constitution].

OBJECTIVE: To investigate the changes of endocrine, cyclic nucleotide and immune systems in subjects of yang deficiency constitution, and to explore the relationship among characteristics and causes of yang deficiency constitution, the physiological and biochemical parameters. METHODS: Based on the diagnostic criteria for the clinical epidemiological investigation, sixty subjects of yang deficiency constitution and fifty of normal constitution were selected. Eight milliliters venous blood were taken from overnight fasted subjects at 8:00-9:00. The sera were obtained by centrifugation of the blood at the speed of 3000 r/min for 5 minutes, and they were stored at -70 degrees C until use. The serum levels of corticosterone, cortisol, adrenocorticotrophic hormone (ACTH), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), free triiodothyronine (FT3), free thyroxine (FT4), thyrotropic-stimulating hormone (TSH), interleukin-1beta and interleukin-2 were measured by enzyme-linked immunosorbent assay; the cAMP/cGMP ratio was also computed; and the differences of the above indexes were compared between the two types of subjects. RESULTS: The serum levels of corticosterone, interleukin-1beta, TSH and cAMP/cGMP ratio of yang deficiency constitution significantly increased as compared with those of normal constitution, and the serum levels of cortisol, ACTH, cGMP and FT4 of yang deficiency constitution significantly decreased in comparison with those of normal constitution. CONCLUSION: Subjects of yang deficiency constitution may be not only related to hypothalamic-pituitary-adrenal axis and hypothalamic-pituitary-thyroid axis, but also related to the functional disorders of cyclic nucleotide and immune systems.

[Mechanism of Yiqi Huayu Bushen Recipe in treating cervical syndrome with kidney deficiency in rats].

OBJECTIVE: To research the action mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound of traditional Chinese herbal medicine, in treating cervical syndrome (CS) with kidney deficiency in rats. METHODS: A total of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, CS with kidney deficiency model group (untreated group) and YHBR group, with 10 rats in each group. Rats in the normal control group received no treatment, while rats of the other two groups underwent resection of both cervical muscles and ovaries to establish the model of CS with kidney deficiency. Three months after surgery, rats in the YHBR group were intragastrically administered YHBR for one month. Another one month later, all rats were sacrificed. The content of serum estradiol (E2) was detected by radio-immunoassay; contents of plasma cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were tested, and the ratio of cAMP/cGMP was also calculated. Hemorheology was detected by Weissenberg's method; expression of alpha-granular membrane protein (CD62p) was detected by flow cytometry; HE staining was used to detect the histopathology of cervical intervertebral disc degeneration; type II collagen protein was detected by immunohistochemistry and aggrecan-1 (Agc1), type II procollagen gene, Col2a1, tissue inhibitor of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-13 (MMP-13) mRNAs were detected by fluorescent quantization polymerase chain reaction. RESULTS: Compared with the untreated group, rats in the YHBR group showed an obvious increase in serum E2 content (P<0.05), and an increase in plasma cAMP and cGMP content without significant difference; hemorheological parameters and percentage of CD62p expression were significantly decreased in the YHBR group (P<0.05, P<0.01). YHBR could improve the degeneration of cervical intervertebral discs, decrease the Miyamoto scores (P<0.05), and increase the type II collagen. The expressions of Agc1, Col2a1 and TIMP-1 mRNAs were significantly increased and MMP-13 mRNA significantly decreased in YHBR group (P<0.05, P<0.01). CONCLUSION: YHBR may improve CS with kidney deficiency and delay the degeneration of cervical intervertebral disc by regulating the immune-metabolism system, coagulation system and endocrine system.

Reduction of atherosclerotic nanoplaque formation and size by Ginkgo biloba (EGb 761) in cardiovascular high-risk patients.

Coating a silica surface with the isolated lipoprotein receptor proteoheparan sulfate (HS-PG) from arterial endothelium and vascular matrices and adding both the atherogenic VLDL/IDL/LDL lipid fraction in its native composition and Ca(2+) ions, we could observe in vitro the earliest stages of atherosclerotic plaque development by ellipsometric techniques (patent EP 0 946 876). This so-called nanoplaque formation is represented by the ternary aggregational complex of the HS-PG receptor, lipoprotein particles and calcium ions. The model was validated in several clinical studies on statins in cardiovascular high-risk patients. In eight patients who had undergone an aortocoronary bypass operation, the reduction of atherosclerotic nanoplaque formation amounted to 11.9+/-2.5% (p<0.0078) and of nanoplaque size to 24.4+/-8.1% (p<0.0234), respectively, after a 2-month therapy with Ginkgo biloba extract (2x 120 mg daily, EGb 761). Additionally, superoxide dismutase (SOD) activity was upregulated by 15.7+/-7.0% (p<0.0391), the quotient oxLDL/LDL lowered by 17.0+/-5.5% (p<0.0234) and lipoprotein(a) concentration decreased by 23.4+/-7.9% (p<0.0234) in the patients' blood. The concentration of the vasodilating substances cAMP and cGMP was augmented by 37.5+/-9.1% (p<0.0078) and 27.7+/-8.3% (p<0.0156), respectively. A multiple regression analysis between the patients' VLDL/IDL/LDL lipoprotein fraction applied in the ellipsometry measurements as well as the further risk factors oxLDL/LDL and Lp(a) on the one hand and changes in nanoplaque formation on the other hand reveals a basis for a mechanistic explanation of nanoplaque reduction under ginkgo treatment. The atherosclerosis inhibiting effect is possibly due to an upregulation in the body's own radical scavenging enzymes and an attenuation of the risk factors oxLDL/LDL and Lp(a).