Effect of chinese herbal drug-containing serum for activating-blood and dispelling-toxin on ox-LDL-induced inflammatory factors’ expression in endothelial cells

<正>Objective:To investigate the effects of drug-containing serum of Chinese herbal compound, Xiongshao Capsule(芎芍胶囊,XS,for activating-blood) and Huanglian Capsule(黄连胶囊,HL,for dispellingtoxin) on the oxidized low-density lipoprotein(ox-LDL)-induced inflammatory factors in human umbilical vein endothelial cells(HUVECs).Methods:Thirty-two rats were randomly divided into four groups:the blank control group treated with distilled water,the positive control group treated with simvastatin(1.8 mg/kg),the test group I treated with Chinese herbal compound of XS(0.135 g/kg),and the test group II treated with Chinese herbal compound of XS(0.135 g/kg) and HL(0.135 g/kg).All the treatments were administered for 7 successive days by gastrogavage.Rats' blood serum was harvested 1 h after the last administration to prepare respective drugcontaining serum.HUVECs were exposed to ox-LDL(100μg/mL) to induce cell injury model and incubated with corresponding drug-containing serum for 24 h.Untreated HUVECs were set for blank control.Levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and soluble intercellular adhesion molecule-1(slCAM-1) in supernatant of cultured HUVECs were determined by enzyme-linked immunosorbent assay(ELISA).HUVEC surface expressions of ICAM-1 and E-selectin were determined by flow cytometry.Results:Levels of IL-6, TNF-α,and slCAM-1 in the supernatant of HUVECs as well as the cell surface expressions of ICAM-1 and E-selectin significantly increased after 24-h ox-LDL stimulation(P0.01),while the abnormal elevations,except slCAM-1 in the test groupⅠ,were all reduced in the treated groups(the positive control and the two test groups) significantly(P0.01 or P0.05).Besides,the effect in the test group II seemed somewhat higher than that in the test group I but with no statistical significance(P0.05).Conclusion:Drug-containing serum of XS plus HL has a certain inhibitory effect on the vascular endothelial inflammation response induced by ox-LDL.     

Serum low-density lipoprotein and heart rate related to the effectiveness in the treatment of angina pectoris with two similar Chinese medicine herbal formulae

Objective  

To explore the effectiveness related indicators which might help identify the indications of Tongxinluo Capsule (通心络胶囊) and Kangxin Capsule (康欣胶囊) targeting on qi deficiency and blood stasis pattern in Chinese medicine (CM) in the treatment of angina pectoris.     

A multi-center randomized double-blind placebo-controlled trial of Xiongshao Capsule (芎芍胶囊) in preventing restenosis after percutaneous coronary intervention: A subgroup analysis of senile patients

Objective  

To evaluate the safety and efficacy of Xiongshao Capsule (芎芍胶囊, XS), consisting of Chuangxiongol and paeoniflorin, in preventing restenosis after percutaneous coronary intervention (PCI) in senile coronary heart disease (CHD) patients.     

Effects of active components of Red Paeonia and Rhizoma chuanxiong on angiogenesis in atherosclerosis plaque in rabbits

Objective： To investigate the effects and mechanism of the active components of Red Paeonia and Fthizoma chuanxiong （Xiongshao Capsule, 芎芍胶囊, XSC） on angiogenesis in atherosclerosis plaque in rabbits. Metbods： Fifty New Zealand rabbits were randomly divided into the normal group, the model group, and the three medicated groups treated respectively with Simvastatin （2.5 mg/kg per day）, low-dose （0.24 g/kg per day） and high-dose （0.48 g/kg per day） XSC, 10 in each group. Rabbits in the normal group were fed with regular diet. To those in the other four groups, high fat diet was given, and a balloon angioplasty was performed two weeks later to establish abdominal aortic atherosclerosis model. Then, the model rabbits were fed continuously with high fat diet, and to those in the medicated groups, the testing drugs were added in the forage correspondingly for 6 successive weeks. Levels of blood lipids were measured at the end of the experiment. Meantime, serum levels of high sensitivity C-reactive protein （hsCRP） and tumor necrosis factor α （TNF-α） were detected with enzyme-linked immunoassay; the plaque area （PA）, cross-sectional vascular area （CVA） and correcting plaque area （PNCVA） were determined quantitatively using imaging software; and the protein expression of vascular endothelial growth factor （VEGF） and factor Ⅷ related antigen （FⅧRAg） in plaque was detected using immunohistochemical method. Results： As compared with the model group, the content of total cholesterol （TC） in the three medicated groups, and contents of triglyceride （TG） and low-density lipoprotein cholesterol （LDL-C） in the Simvastatin group were lower to various extents （P〈0.05, P〈0.01）. The serum level of hsCRP in all modeled rabbits was higher than that in the normal group, but in the three treated groups it was significantly lower than that in the model group （P〈0.05, P〈0.01）. Expressions of VEGF and FⅧRAg, as well as PNCVA in the three medicated groups were significantly lower than those in the model control group （P〈0.05, P〈0.01）. Conclusion： The active components of Red Paeonia and Rhizoma chuanxiong have definite effects in delaying the genesis and development of atherosclerosis, its mechanism might be related with the inhibition on angiogenesis in plaque, and also with its actions of lipo-metabolism regulation and anti-inflammation.     

Xiongshao Capsule (芎芍胶囊) promotes angiogenesis of HUVEC via enhancing cell proliferation and up-regulating the expression of bFGF and VEGF

Objective  

To evaluate the angiogenic effect of the Xiongshao capsule (芎芍胶囊, XSC) in human umbilical vein endothelial cells (HUVEC), and to investigate the possible molecular mechanisms mediating its biological effect.     

Effect of Xuefu Zhuyu Capsule (血府逐瘀胶囊) on the Symptoms and Signs and Health-related Quality of Life in the Unstable Angina Patients with Blood-stasis Syndrome after Percutaneous Coronary Intervention: A Randomized Controlled Trial

Objective: Compared with Shengmai Capsule (生脉胶囊, SM), the study was conducted to evaluate the efficacy and safety of Xuefu Zhuyu Capsule (血府逐瘀胶囊, XFZY) on the symptoms and signs and health-related quality of life (HR-QOL) in the unstable angina (UA) patients with blood-stasis syndrome (BSS) after percutaneous coronary intervention (PCI). Methods: A randomized, double-blinded, doubledummy, and placebo-controlled trial was applied. Ninety patients, diagnosed as UA and BSS after successful PCI, were enrolled and equally randomized into three groups, XFZY group, SM group, and placebo group, and administered with the corresponding medications respectively for four weeks. The clinical symptoms and signs (CSS), electrocardiography (ECG), and BSS scores were recorded and compared among groups during and after the treatment. Short-form 36 (SF-36) and Seattle Angina Questionnaire (SAQ) were applied to assess the HR-QOL in each group before and after the treatment. Safety indexes (blood routine and liver and kidney function tests) were also examined at the beginning and after the treatment. Results: Eighty-six patients completed the whole study. After the treatment, the total effective rates of the XFZY group in ameliorating CSS and ECG were 76.7% and 60.0%, respectively, which were obviously higher than those in SM (CSS: 53.3%; ECG: 36.7%) and the placebo (CSS: 43.3%; ECG: 30.0%) groups. After one week's treatment, BSS scores slightly decreased in each group, but no significant differences were found among three groups (P0.05). After four weeks' treatment, BSS scores in the XFZY group decreased to a lower level compared with SM (P0.05) and the placebo (P0.01) groups. After the treatment, the efficacy of XFZY group in improving body pain (BP), general health (GH), vitality (VT), society functioning (SF), role emotional (RE), angina stability (AS), angina frequency (AF), and treatment satisfaction (TS) were better than those in the placebo group (P0.05, P0.01). Meanwhile, the dimensions of BP, GH, SF, AS, AF, and TS were better improved than those in the SM group (P0.05). No obvious adverse reaction was found during and after the treatment except one case in the XFZY group reporting of stomach discomfort. Conclusions: Compared with SM Capsule treatment, a short-term treatment with XFZY Capsule exhibits better efficacy on CSS and BSS scores, and HR-QOL in UA patients with BSS after PCI. However, its long-term efficacy and safety still needs further investigation.     

Effect of Feitai Capsule(肺泰胶囊) on Quality of Life and Progression-Free Survival of Patients with Unresectable Non-Small Cell Lung Cancer

<正>Objective:To examine the effect of a Chinese medicinal herbal formula(Feitai Capsule,肺泰胶囊) on the quality of life(QOL) and progression-free survival(PFS) of patients with unresectable non-small cell lung cancer(NSCLC).Methods:Sixty-two patients were randomly divided into the treatment group(31 cases) and the control group(31 cases).For the treatment group,4 capsules(1.2 g/capsule) of Feitai Capsule were administered 3 times a day after meals for 3 weeks;then no drug was administered for 1 week.This schedule was continued for at least 3 more cycles(12 weeks totally).If there were no obvious toxic reactions,the treatment was extended.The patients were evaluated at least once every 8 weeks until progressive disease(PD). For the control group,the regular follow-up and evaluation were performed at least once every 8 weeks until PD.Clinical symptoms,objective response,physical constitution and energy,QOL,and PFS were evaluated regularly.Analysis of variance(ANOVA),a non-parametric test,and analysis of covariance were used to compare clinical features,amelioration of clinical symptoms,physical constitution and energy,and QOL.Kaplan-Meier analysis was used to compare the two-group PFS.Results:Sixty patients finished the final evaluation, with 30 patients in each group.Baseline characters between groups were not significantly different(P0.05). The control group had a 36.7%improvement in clinical symptoms,while the treatment group had a 73.3% improvement.This difference was statistically significant(Z=-2.632,P=0.008).The control group had a 26.7% improvement in the Karnofsky performance status(KPS),while the treatment group had a 53.4%improvement. This was also significantly different(Z=-2.182,P=0.029).A comparative analysis indicated a positive correlation (r=0.917,P0.001).Compared with the control group,QOL in the treatment group was significantly improved, except in the social/family condition and doctor-patient relationship indicators.The PFS of the treatment group and control group were 6.23 months and 4.67 months,respectively(P=0.048).Conclusion:Feitai Capsule,a Chinese medicinal herbal treatment could improve the QOL and extend the PFS of the unresectable NSCLC patients.     

Efficacy of dual antiplatelet therapy combined with Naoxintong Capsules (脑心通胶囊) following coronary microembolization induced by homologous microthrombi in rats

Objective  

To evaluate the effificacy of dual antiplatelet therapy combined with Naoxintong Capsule (脑心通胶囊, NXTC) in a rat model of coronary microembolization (CME).     

Effect of Xinfeng Capsule (新风胶囊) on the cardiac function in patients with rheumatoid arthritis

Objective  

To study the changes in cardiac function of rheumatoid arthritis (RA) patients and: to observe the effect of Xinfeng Capsule (新风胶囊, XFC) on them.     

Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by Tougu Xiaotong Capsule (透骨消痛胶囊)-containing serum

Objective  

To study the mechanism of action of Tougu Xiaotong Capsule (透骨消痛胶囊, TGXTC) ex vivo in suppressing chondrocyte (CD) apoptosis induced by sodium nitroprussiate (SNP).     

Effects of Tongguan Capsule(通冠胶囊)on Post-myocardial Infarction Ventricular Remodeling and Cardiac Function in Rats

<正>Objective:To observe the effects of Tongguan Capsule(通冠胶囊,TGC) on post-myocardial infarction ventricular remodeling and heart function in rats.Methods:A rat model of acute myocardial infarction (AMI) was established by coronary ligation.Experimental rats were randomized to 4 groups including three model groups(Group A:captopril 5 mg/kg·day,n=7;Group B:TGC 10 g/kg·day,n=7;and Group C:placebo, n=8),and a sham-control group(Group D:blank control,n=6).Animals were treated for 4 weeks.The cardiac function of rats was assessed at the end of the experiment based on left ventricular ejection fraction(LVEF) and left ventricular short axis fractional shortening(LVFS) detected by colored echocardiography;meanwhile, the condition of ventricular remodeling was observed through the levels of left ventricular mass(LVM),plasma aldosterone(ALD),myocardial angiotensinⅡ(AngⅡ) and myocardial collagen measurements.Results:At the end of the experiment,LVEF and LVFS in Group A and B were improved significantly,while those in Group C were unchanged,the LVEF in Group A,B,C,and D was 0.57±0.46,0.61±0.08,0.36±0.55 and 0.76±0.02,respectively;and their LVFS was 0.31±0.52,0.34±0.04,0.23±0.57 and 0.45±0.03,respectively.The difference was statistically significant when comparing the two indexes in Group A and B with those in Group C and D(P0.05).LVM,levels of plasma ALD and myocardial AngⅡwere lower in Group A and B than in Group C,but a comparison between Group A and B showed an insignificant difference in lowering LVM and ALD,while the lowering of AngⅡwas more significant in Group B than in Group A(754.7±18.7 pg/mL vs 952.6±17.6 pg/ mL,P0.05).Morphological examination showed that in Group A and B the swollen myocardial cells had shrunk, with regularly arranged myocardial fibers and decreased collagen proliferation,but the improvements in Group B were more significant.Conclusion:TGC could markedly improve the post-infarction ventricular remodeling and cardiac function in rats,showing that the efficacy was better than or equal to that of captopril.     

Effect of Xuezhikang Capsule(血脂康胶囊)on Serum Tumor Necrosis Factor-αand Interleukin-6 in Patients with Nonalcoholic Fatty Liver Disease and Hyperlipidemia

<正>Objective:To evaluate the effect of Xuezhikang Capsule(血脂康胶囊) on the serum levels of inflammatory factors such as tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in patients with nonalcoholic fatty liver disease(NAFLD) and hyperlipidemia,and to explore whether it has anti-inflammatory effect.Methods:A total of 84 patients were randomly assigned to two groups with stratified block randomization, the treatment group(42 cases) and the control group(42 cases).They were treated with Xuezhikang Capsule and polyene phosphatidylcholine capsule for twenty-four weeks,respectively.The changes in serum TNF-αand IL-6 were measured by enzyme linked immunosorbent assay before treatment and at the 12th and 24th week. Results:Compared with those before treatment,the serum levels of TNF-αand IL-6 significantly decreased in both groups after treatment(P0.01).There was no significant change between the two groups for the treatments at different time points(P0.05) and between the two groups for treatments at the same time points (P0.05).Conclusion:Xuezhikang Capsule can inhibit the serum inflammatory factor in patients with NAFLD and hyperlipidemia.     

Effect of Chinese Drugs for Activating Blood Circulation and Detoxifying on Indices of Thrombosis Inflammatory Reaction and Tissue Damage in A Rabbit Model of Toxin-Heat and Blood Stasis Syndrome

Objective:To observe the regulatory effect of Chinese drugs for activating blood circulation(ABC) and for activating blood circulation and detoxifying(ABCD) on indices of thrombosis,inflammatory reaction, and tissue damage in a rabbit model of toxin-heat and blood stasis syndrome.Methods:Fifty-four rabbits were randomized into the normal control group,model group,simvastatin group(simvastatin,0.93 mg/kg per day),ABC group[Xiongshao Capsule(芎芍胶囊),0.07 g/kg per day],and ABCD group[Xiongshao Capsule,0.07 g/kg per day,and Huanglian Capsule(黄军胶囊),0.14 g/kg per day].All except the normal control group received a single injection of bovine serum albumin and were fed with high-fat diets for 6 weeks.At the end of week 4 of giving highfat diets,a dose of endoxitin was given by ear vein injection,and a randomized 2-week treatment was initiated. At the end of treatment,blood lipids,circulating endothelial cells,and the pathological changes of the aortic arch were assessed.The serum levels of matrix met alloproteinases(MMP-9),tissue inhibitors to met alloproteinase (TIMP-1),granule membrane protein-140(GMP-140),plasminogen activator inhibitor-1(PAI-1),high-sensitivity C-reactive protein(hs-CRP),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α) were determined.Results: Compared with the model group,ABCD group showed decreased serum triglyceride(TG) level,improvement in the pathological change in the aortic arch,and reduction in the number of circulating endothelial cells(4.00±1.41 per 0.9μL for ABCD group vs 7.83±1.72 per 0.9μl L for the model group).In addition,the levels of serum GMP-140,PAI-1,and IL-6 in ABCD group were also significantly reduced[0.79±0.20 ng/mL,5.23±1.39 ng/mL, 40.64±10.11 pg/mL for ABCD group vs 1.08±0.31 ng/mL,7.28±2.01 ng/mL,54.44±13.56 pg/mL for the model group,respectively,P<0.05].A trend showing improvement in the indices of thrombosis,inflammatory reaction, and tissue damage was observed in the ABC group when compared to the model group,but the changes were not statistically significant(P>0.05).Conclusions:Chinese drugs for activating blood circulation and detoxifying have beneficial effects on regulating indices of thrombosis(GMP-140 and PAI-1) and inflammatory reaction(IL-6) in rabbit model with toxic-heat and blood stasis.The effect of the activating blood circulation and detoxifying drugs in regulating the levels of serum GMP-140,PAI-1,and IL-6 was superior to that of the activating blood circulation drugs.     

Treatment of Non-alcoholic Fatty Liver Disease by Qianggan Capsule(强肝胶囊)

<正>Objective:To observe the therapeutic effect and safety of Qianggan Capsule(强肝胶囊,QGC) in treating non-alcoholic fatty liver disease(NAFLD),using polyene phosphatidylcholine capsule(PPC) as a reference.Methods:Eighty-eight patients with NAFLD were randomly assigned to two groups,45 in the treatment group treated with QGC and 43 in the control group treated with PPC.The course of treatment lasted for 6 months.Changes in liver function,blood lipids,and iconographic indexes before and after treatment were observed,and clinical efficacy was evaluated.Results:In the treatment group,alanine aminotransferase(ALT) was lowered significantly from 56.02±32.59 IU/L before treatment to 38.27±22.68 IU/L after treatment,and CT liver/spleen ratio significantly increased from 0.69±0.18 to 0.91±0.25,showing statistical significance(P0.05);in contrast,the corresponding changes of the two indexes in the control group were 56.56±26.33 IU/L to 49.67±26.22 IU/L, and 0.66±0.20 to 0.75±0.24,respectively,the pre-post treatment difference showing insignificant difference(P0.05). No severe adverse reactions occurred during the whole treatment course.Conclusion:QGC is an effective and safe remedy for the treatment of NAFLD.     

Effects of Combined Therapy of Xuezhikang Capsule(血脂康胶囊)and Valsartan on Hypertensive Left Ventricular Hypertrophy and Heart Rate Turbulence

<正>Objective:To observe the effect of combined therapy with Xuezhikang Capsule(血脂康胶囊,XZK) and Valsartan on left ventricular hypertrophy(LVH) and heart rate turbulence(HRT) in hypertensive patients. Methods:Ninety primary hypertensive patients with LVH were randomly assigned to three groups.Basic treatment,including aspirin,β-blockers,calcium antagonists,etc.were administered to all patients.Additionally, Valsartan(VS,80 mg once a day) was given to the 30 patients in the VS group.Valsartan(in the same dosage) and XZK(600 mg,twice a day) were given to the 32 patients in the Chinese medicine(CM) group,while none was given to the 28 patients in the control group.The therapeutic course lasted for 24 months.Changes in left ventricular mass index(LVMI) measured by cardiac ultrasonic indices,HRT parameters,including the original heart rate(TO) and slope coefficient(TS),systolic and diastolic blood pressures(SBP and DBP),as well as blood cholesterol level(TC) were measured before and after treatment.Results:After treatment,TO and LVMI were lowered,while TS increased in both the VS group and the CM group(P0.01),but changed insignificantly in the control group.Significant differences between the CM group and the control group were shown in terms of TO,LVMI,SBP,DBP and TS(P0.01);and between the CM group and the VS group in terms of TO,LVMI and TS(P0.01).Moreover,HRT parameters showed an evident correlation with LVMI(r=0.519-0.635,P0.01). Conclusion:Combined therapy with XZK and Valsartan can improve hypertensive LVH and HRT parameters, and lessen the damage on the autonomous nervous system.     

Effects of Corocalm (Shuguan Capsule, 疏冠胶囊) on acute myocardial ischemia in anesthetized dogs

Objective:To investigate the effects of Corocalm (Shuguan Capsule,疏冠胶囊) on acute myocardial ischemia in anesthetized dogs and its possible therapeutic mechanism.Methods:The acute ischemia model was established by ligating the left anterior descending (LAD) artery.Twenty- five dogs were randomly divided into 5 groups (5 clogs in each group):the control group (treated with normal saline 3 mL/kg),the refined Guanxin Capsule group (精制冠心胶囊,GXC 200 mg/kg),high and low dose Corocalm groups (48.5 mg/kg for low dose group and 194.0 mg/kg for high dose group) and the Diltiazem group (5 mg/kg).The animals were treated via a single duodenal administration after the model was established.The experiments used epicardial electrocardiogram (EECG) to measure the scope and degree of myocardial ischemia.Simultaneously,the coronary blood flow (CBF) and serum activity levels of creatine phosphokinase (CK) and lactate dehydrogenase (LDH) were measured by electromagnetic flow meter and automatic biochemical analyzer respectively.The plasma endothelin (ET) content was quantified by radioimmunoassay.Results:Corocalm (48.5 mg/kg and 194.0 mg/kg) significantly decreased the degree and scope of myocardial ischemia,reduced the infarct area,markedly increased the CBF,and inhibited the increase of CK and LDH activities and ET levels induced by myocardial ischemia/infarction.Conclusion:Corocalm could improve the state of acute myocardial ischemia and infarction in dogs.The mechanism of action might be correlated to increasing CBF, inhibiting CK and LDH activities and preventing ET release.     

Short-term Effect of Combined Therapy with Jinlong Capsule(金龙胶囊)and Transcatheter Arterial Chemoembolization on Patients with Primary Hepatic Carcinoma and Its Influence on Serum Osteopontin Expression

<正>Objective:To observe the clinical combination effect of Jinlong Capsule(金龙胶囊,JLC) and transcatheter arterial chemoembolization(TACE) on the patients with primary hepatic carcinoma(PHC) and JLC's influence on serum osteopontin(OPN) expression and elucidate the correlation between the serum OPN level and curative effect of JLC and TACE.Methods:A total of 98 patients with PHC were observed in a randomized controlled trial(RCT).They were assigned to the Chinese medicine(CM) group(53 patients who were treated with TACE and JLC) and the intervention group(45 patients who were treated with TACE only).The serum OPN levels were measured before and after treatment by quantitative sandwich enzyme-linked immunosorbent assay(ELISA).Forty healthy people were assigned to the control group.The clinical efficacy was observed and Karnofsky score(KPS) was graded.Results:The clinical efficacy of the CM group(60.38%) was better than that of the intervention group(40.00%),and the KPS(84.35±12.19) was higher than the intervention group(69.86±11.58)(P0.05).The serum OPN levels before and after treatment in the patients with PHC were significantly elevated compared with those in the control group(P0.01).After treatment,the OPN levels in CM group(117.69±78.50) were significantly lower compared with those in intervention group(151.09±83.90,P0.05).The OPN levels of responders were remarkably lowered than the non-responders after treatment,and the level of OPN in the CM group was lower than the intervention group(P0.05).Conclusions:The short-term clinical efficacy and the quality of life of patients with PHC can be improved by combining JLC with TACE.The serum OPN levels in PHC patients can reflect the curative effect of treatment and the prognosis of the disease.     

Effect of Chinese drugs for supplementing qi, Nourishing Yin and activating blood circulation on myocardial perfusion in patients with acute myocardial infarction after revascularization

Objective:To observe the effects of Chinese drugs for supplementing qi,nourishing yin and activating blood circulation on the myocardial perfusion in acute myocardial infarction(AMI) patients after revascularization.Methods:Eighty patients with anterior or inferior ventricular wall AMI,who had received revascularization by intravenous thrombolysis or coronary bypass,were randomized into the treated group and the control group equally,both treated with conventional Western medical treatment,but combined,r...     

The potential role of Chinese medicine in ameliorating extra-articular manifestations of rheumatoid arthritis

Rheumatoid arthritis(RA) is a chronic systemic autoimmune disease chiefly affecting synovial membranes of multiple joints.The clinical manifestations are highly variable.Besides joint affection,extra-articular manifestations always occur in RA patients,such as lung,blood vessel,heart,endocrine glands,hematological system, and nervous system affections.In addition to Western medicine therapy,Chinese medicine also plays a significant role in the treatment of RA with good efficacy and less adverse reactions.This paper summarizes the effects of Xinfeng Capsule(新风胶囊),a Chinese medicine,and the mechanisms of its action in ameliorating the extra-articular manifestations based on a series of clinical and experimental researches.