Large-scale clinical trial is an important measure of clinical evaluation on drugs. This paperintroduces the concept and features of large-scale clinical trial, the possibility and necessity of large-scale clinical trial of traditional Chinese medicine, as well as its administration and quality control, with Myocardial Infarction Secondary Prevention Study in Traditional Chinese Medicine (MISPS-TCM), a National Program Subject, as an example.

Relapse and metastasis are the main causes responsible for failure of surgical operation in treating malignant tumors. This article introduces the present development in post-operation treatment of lung cancer, liver cancer and stomach cancer, showing that the synthetic treatment by stages combined with traditional Chinese medicine is the important way to prevent and treat the post-operation relapse and metastasis of tumors, and suggesting the optimized method for treatment.

There are two stages for selecting the clinical intervention outcome variables. One is crude collection and the other is precise collection. The selection methods include brainstorming method, Delphi method, Gordon method, systematic review, systematic analysis method, mathematic model method and so on. Each method has some advantages or disadvantages. When selecting the clinical intervention outcome variables, researchers should make sure the research aim, the representative and the sensitivity of the variables, and select all kinds of recommended methods comprehensively, then decide which method should be used.

Objective: To evaluate the safety and efficacy of Yiqi Pingchuan Granule in treating acute attack of asthma due to qi deficiency and cold syndrome. 

Methods: A randomized controlled trial was conducted. A total of 80 patients with an acute attack of asthma were included. They were allocated into two groups randomly in a ratio of three to one. Sixty patients in the treatment group received Yiqi Pingchuan Granule and 20 patients in the control group received Ruyi Dingchuan Pill. Patients in both groups were treated for 7 days.

Results: There were no significant differences in traditional Chinese medicine syndrome, clinical symptoms and lung function between the two groups (P>0.05). After treatment, the forced expiratory volume in 1 second was increased in the treatment group (P＜0.05), and the peak expiratory flow was accelerated in the control group (P＜0.05). No significant side effects were noted in both groups. 

Conclusion: Yiqi Pingchuan Granule is safe and effective in treating acute attack of asthma due to qi deficiency and cold syndrome.

Objective: To study the effects of Salvia miltiorrhiza on serum levels of cytokines including interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) in patients with severe acute pancreatitis (SAP). 

Methods: Thirty-six SAP patients were randomly divided into Salvia miltiorrhiza-treated group and non-Salvia miltiorrhiza-treated group with eighteen patients in each group. Fourteen age-matched healthy volunteers were assigned to control group. Serum levels of IL-6, IL-8, and TNF-α were tested within 24 hours of admission and 7 days after admission. 

Results: Compared with those of the healthy volunteers, serum levels of the three cytokines of the SAP patients were significantly elevated (P＜0.05). Serum levels of the three cytokines were significantly decreased in both Salvia miltiorrhiza-treated group and non-Salvia miltiorrhiza-treated group after 7-day treatment (P＜0.05). The declining of the serum levels of the three cytokines in Salvia miltiorrhiza-treated group was more obvious than that in non-Salvia miltiorrhiza-treated group.

Conclusion: Salvia miltiorrhiza can treat SAP patients by reducing the serum levels of IL-6, IL-8, and TNF-α.

Objective: To make an animal model of cervical spondylosis (arthralgia syndrome type) with stimulation of wind, cold, and dampness. 

Methods: Twenty-four 8 months old male New Zealand white rabbits were randomly allocated into four groups: normal control group, light stimulation group, moderate stimulation group and severe stimulation group. The wind speed was 10.8-13.8 m/s, the temperature was (5±0.5) ℃, and the humidity was 100%. The rabbits of light, moderate, and severe stimulation groups were kept in the above-mentioned environments for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively. The intervertebral discs were stained with HE method, and observed with a light microscope. Prostaglandin E₂ (PGE₂), 6-ketone-prostaglandin F_1α (6-K-PGF_1α) and thromboxane B₂ (TXB₂) contents were measured by ELISA. Fas and Bcl-2 expressions were examined by immunohistochemical avidin-biotin peroxidose complex technique. Interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α), and transforming growth factor β (TGF-β) mRNA expressions were examined by reverse transcription-polymerase chain reaction.

Results: The nucleus pulposus of rabbits in the light and moderate stimulation groups shrunken, and in the severe stimulation group, the anulus fibrosus loosed or ruptured, and the cartilage end-plate became proliferated. Compared with rabbits in the normal control group, the PGE₂ content rose in the light stimulation group, the contents of PGE₂, 6-K-PGF_1α, and TXB₂ increased, the expressions of IL-1β and TNF-α mRNAs and Fas were up-regulated, and the expressions of TGF-β mRNA and Bcl-2 were down-regulated in the moderate and severe stimulation groups. The expression of Fas was up-regulated mostly and Bcl-2 was down-regulated mostly in the severe group. 

Conclusion: Moderate and severe stimulations of wind, cold and dampness can lead to degeneration of cervical intervertebral discs of rabbits. The model corresponds to the theory of traditional Chinese medicine about arthralgia syndrome caused by wind, cold and dampness.

Objective: To investigate whether the water extractives of regulating qi and blood prescription (WQBP) had effects on early atherosclerosis of apolipoprotein E-deficient mice (ApoE^- mice) at the age of 19 weeks or not, and to explore the possible mechanisms. 

Methods: Forty ApoE^- mice, six weeks of age, were given high-fat diet and randomly divided into four groups: high-dose WQBP-treated group (360 mg/kg), low-dose WQBP-treated group (72 mg/kg), simvastatin-treated group (25 mg/kg) and untreated group, with ten mice in each group. Meanwhile, ten C57BL/6 mice of same genetic background were allocated to normal control group. Mice in the high- and low-dose WQBP-treated groups and simvastatin-treated group were administered with corresponding drugs from the 15 to 19 weeks. Mice in the untreated and normal control groups were administered with isovolumic water. Sacrificed at 19 weeks, the level of blood-lipid, the plaque construction, plaque integral, and the contents of plaque macrophages and vessel smooth muscle cells of the mice were analyzed by immunohistochemical method and a computer picture processing system.

Results: Compared to the untreated group, high-dose WQBP group could obviously decrease the level of low-density lipoprotein cholesterol (LDL-C). Simvastatin group could decrease the levels of LDL-C and total cholesterol (TC) (P＜0.01). In high-dose WQBP-treated group and simvastatin-treated group, the thickness of fiber cap and the quantities of vessel smooth muscle cells increased (P＜0.05), the quantities of plaque macrophages and the ratio of lipid and plaque reduced (P＜0.01). 

Conclusion: WQBP and simvastatin can interfere in early atherosclerosis of ApoE^- mice, attenuate and stabilize plaque in some extent. The mechanisms may include adjusting blood lipid, decreasing macrophage number and increasing the quantities of vessel smooth muscle cells.

Objective: To explore the effects of Gymnadenia conopsea alcohol extract (GcAE) on the collagen synthesis in rat lungs exposed to silica and the influence on antioxidase activities, level of lipid peroxidation (LPO).

Methods: One hundred and twenty rats were randomly divided into control group, silica group, and GcAE-treated group. Silicotic animal models were established by direct tracheal instillation of silica into rat lungs surgically. From the second day of model establishment, rats in GcAE-treated group were orally given GcAE [8 g/（kg·d） corresponding to raw herb]. At 7, 14, 21, 28 and 60 days after establishment of the animal model, eight rats in each group were sacrificed, and samples were collected. The malondialdehyde (MDA) content, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in plasma were assayed by a spectrophotometer. Types Ⅰ and Ⅲ collagen were detected by Sirius red polarization and microscopy, and measuered by Image-Pro Plus Version 4.5 for Windows software.

Results: GcAE could reduce the lung/body weight ratio of rats exposed to silica, the synthesis of types ⅠandⅢcollagen of the lungs and the level of lipid peroxidation, increase the activities of SOD and GPx.

Conclusion: GcAE can ameliorate the silica-induced pulmonary fibrosis by increasing the activities of antioxidase and alleviating the damage of lipid peroxidation to the lungs.

Objective: To screen antitumor active compounds, drug-like or leading compounds from Chinese traditional and herbal drugs.

Methods: Eleven coumarin compounds isolated from the Chinese traditional and herbal drugs were studied for their antitumor activities in vitro by determining the inhibition rates against growth of human bladder carcinoma cell line E-J.

Results: It showed that umbelliferone, scoparone, demethylfuropinarine, isopimpinellin, forbesoside, columbianadin, decursin and glycycoumarin inhibited the growth of human bladder carcinoma cell line E-J in vitro and their activities showed a concentration-effect relationship. The inhibitory effects of forbesoside, columbianadin, decursin and umbelliferone, with IC50 values of 7.50×10 ^-7, 2.30×10 ^-6, 6.00×10 ^-6 and 1.30×10 ^-6 mol/L, respectively, were stronger than those of the other tested compounds. However, xanthotoxin, esculin and sphondin did not inhibit the growth of human bladder carcinoma cell line E-J in this assay condition. 

Conclusion: These findings indicate that forbesoside, columbianadin, esculin, decursin and umbelliferone would be effective or regarded as potent drug-like or leading compounds against human bladder carcinoma.

Objective: To investigate the effects of salvianolic acid B (SA-B) on portal hypertension induced by endothelin-1 in rats. Methods: Twenty-eight Sprague-Dawley rats were randomly divided into four groups: ET-1 group, ET-1+SA-B group, ET-1+ETAR blocker (BQ-123) group and ET-1+ETBR blocker (BQ-788) group. The rats of ET-1+SA-B group underwent intragastrical administration of salvianolic acid B for five days before ET-1 injection, while in three other groups' drinking water was given. In BQ-123 group or BQ-788 group, an intravenous injection of BQ-123 or BQ-788 via femoral vein was administered 30 minutes prior to ET-1 injection. Then changes of portal pressure, cervical artery pressure and heart rate were monitored continuously. Results: After ET-1 injection, the portal pressure of all rats in the ET-1 group increased significantly, while slightly in groups that pretreated with SA-B, BQ-123 or BQ-788. Conclusion: SA-B can attenuate the elevated portal pressure induced by ET-1 with effect similar to ETR blocker.

Objective: To explore the effects of inactivated rabbit serum containing compound realgar and natural indigo tablet (CRNIT) on cell line NB4. 

Methods: The experimental rabbits were taken as the provider of the animal serum, and the serum was inactivated before the experiment. The serum was divided into two groups based on whether the rabbits were given CRNIT. The concentration of arsenic in the rabbit's serum was detected by AFS-230a double path atom fluorescence photometer. The inhibition rates and apoptosis rates were regarded as the observational indexes.

Results: The concentration of arsenic in the inactivated rabbit serum containing and not containing the drug were (0.010 0±0.001 0) mg/L and (0.110 0±0.006 4) mg/L respectively, and the difference had statistical significance (P＜0.01). The two groups of serum all had inhibitory effect on the growth of NB4 cells depending on the drug concentration and effect time. And there were significant differences among the groups. The two groups of serum all induced the apoptosis of NB4 with positive relations with the concentration and effect time. And there were significant differences among the groups.

Conclusion: The rabbit serum containing CRNIT can obviously restrain the growth of NB4 cells and the inhibitory effect depends on the concentration and effect time. And the rabbit serum containing CRNIT can also induce the apoptosis of NB4 cell line and the apoptosis rates depend on the concentration and effect time.

Objective: To study the mechanism of herbal application along meridians for treatment and prevention of asthma by using serum pharmacological test to observe the effects of serum containing herbs against the constriction of tracheal spiral strips induced by acetylcholine chloride (Ach).

Methods: Guinea pigs were randomly divided into normal control group, normal saline (NS) application group, aminophylline application group, aminophylline injection group, 1-day herb application group, 7-day herb application group and 14-day herb application group. Asthma was induced by Hutson's method in guinea pigs except the normal control group. Guinea pigs in herb application groups were treated by external application of a compound herbal medicine 60 min once every day. Guinea pigs in NS application group were treated by external application of NS. Guinea pigs in the two aminophylline-treated groups were treated by external application and intraperitoneal injection of aminophylline at a dose of 400 mg/kg, respectively. The guinea pigs were killed and the sera were obtained after 1-day, 7-day and 14-day treatment in the three herb application groups, 7-day treatment in the NS application group, the aminophylline application and injection groups, respectively. Serum pharmacological method was used to do the experiment, the effects of different sera on the constriction of tracheal strips were observed, and the constriction rates were calculated.

Result: The serum containing herbs had an effect in reducing the constriction of tracheal spiral strips induced by Ach, and the effect was similar to that of the serum obtained from the aminophylline injection group. The constriction rate of the tracheal spiral strips was decreased when herbal application treatment was prolonged within a period of time, and it became stable when herbal application treatment was between 7-14 days. The constriction of tracheal spiral strips induced by Ach could be reduced remarkably when it was previously treated by serum containing herbs. 

Conclusion: The anti-acetylcholine function with a time-dependent effect is one of the mechanisms of herbal application treatment along meridians for asthma, and furthermore, herbal application treatment along meridians might be useful for preventing asthma.

Objective: To compare the acute toxicity of common injection and sustained-release preparation of norcantharidin for mice so as to identify the attenuation of the sustained-release preparation of norcantharidin. 

Methods: The poloxamer 407 was used as a sustained-release vehicle for topical administration of norcantharidin, and the acute toxicity of mice treated with common injection and sustained-release preparation of norcantharidin was observed. The median lethal dose (LD₅₀) was calculated by Bliss software. 

Results: The symptoms of mice were similar between the two groups, but the appearance of symptoms in norcantharidin/poloxamer 407 group was 4 hours later than that in norcantharidin group. The LD₅₀ of norcantharidin administered through vein injection was 12.6 mg/kg. The LD₅₀ of norcantharidin/poloxamer 407 administered through intraperitoneal injection, intrahepatic injection and intramuscular injection were 19.9, 19.1 and 20.9 mg/kg, respectively, and the LD₅₀ of the common preparation were 13.0, 13.1 and 15.1 mg/kg, respectively. 

Conclusion: The norcantharidin/poloxamer 407 is less toxic than the equivalent dose of norcantharidin, mainly because norcantharidin/poloxamer 407 may release norcantharidin sustainedly, thus reducing norcantharidin concentration in blood.

Objective: To observe whether injection of medicine into low hydraulic resistance point along meridian brings about higher medicinal effect and to explore the efficacy of the theory that meridians are made up of channels featuring low hydraulic resistance by observing the diuretic effect of injecting furosemide or saline into the low hydraulic resistance point Shuifen (CV 9), vein and Zusanli (St 36) respectively. 

Methods: Acute edema was induced in pigs by rapid intravenous injection of 2 000 ml normal saline. The pigs were divided into four groups: Shuifen (CV 9) injection of half dose furosemide group (SFF group), intravenous injection of full dose furosemide group (VF group), Zusanli (St 36) injection of full dose furosemide group (ZSLF group), and Shuifen (CV 9) injection of half dose normal saline group (SFS group). The accumulated urine quantity and the urine quantity generated in every 15-minute period were measured in each group respectively, every 15 minutes after injection, and the measurement lasted for two hours at one experiment. Each group involved eight times of experiments with one pig used for one experiment, which means the whole observation involved 32 times of experiments. 

Results: The accumulated urine quantities observed in both SFF group and VF group were higher than those in the ZSLF group and the SFS group all through the measurement, showing significant differences during the period from the 15th minute to the 45th minute (P＜0.05). But no significant difference was observed between the SFF group and the VF group during the whole 2-hour measurement (P>0.05). Analysis of urine quantity generated in every 15-minute period showed that diuretic effect climaxed during the 15th minute to the 30th minute in both SFF group and VF group. By contrast, ZSLF group reached diuresis climax during the 45th minute to 60th minute and no diuresis climax was observed in the SFS group all through the measurement.

Conclusion: Injection of medicine into low hydraulic resistance point along meridian generates faster and more powerful medicinal potency, and this is likely to be applied to clinical practice. The theory that meridians are channels featuring low hydraulic resistance is important to the elucidation of meridians.

Objective: To introduce the method of analyzing repeated data measured by water maze with SPSS 11.0, and offer a reference statistical method to clinical and basic medicine researchers who take the design of repeated measures. 

Methods: Using repeated measures and multivariate analysis of variance (ANOVA) process of the general linear model in SPSS and giving comparison among different groups and different measure time pairwise. 

Results: Firstly, Mauchly's test of sphericity should be used to judge whether there were relations among the repeatedly measured data. If any (P≤0.05), multivariate ANOVA should be taken next, or Greenhouse-Geisser corrected results should be taken. Treated effect could be evaluated by estimating between-subject variance. Repeated measurement effect or its interactive effect with treated group could be evaluated by estimating within-subject variance. The method of Bonferroni should be used to do pairwise comparisons of the repeatedly measured data in different measurement time of each treated group. With multivariate ANOVA, data in different treated group of each measurement time could be compared pairwise. 

Conclusion: The repeated measures process of the general linear model is suitable for variance analysis of repeatedly measured data. SPSS statistical package is available to fulfil this process.