OBJECTIVE: To investigate the separation and purification technology of total saponins from the root of Thladiantha dubia (TSTR). METHODS: The content of TSTR was determined by UV-visible spectrophotometry. By comparing static adsorption and desorption properties of different types (AB-8, D101, DM130, HPD100, HPD300, HPD450, HPD600, HPD826, NKA-9) of macroporous adsorption resin, the type of macroporous adsorption resin was screened. With the content of TSTR as the index, influential factors of macroporous adsorption resin for adsorbing (ratio of height to diameter of resin, mass concentration of medicine liquid, adsorption volume flow, saturated extent of adsorption) and desorbing (desorption solvent volume fraction, desorption solvent volume flow, volume of desorbed solvent) TSTR were investigated. The optimal technology was screened. The technology validation, purification and preparation were conducted. RESULTS: HPD100 type macroporous adsorption resin had good adsorption and desorption properties for TSTR. The optimal adsorption technology was that the ratio of the height to diameter of the resin column was 1:5; mass concentration of medicine liquid was 1 g/mL; adsorption volume flow rate was 1 BV/h; saturated adsorption capacity was 1. 25 g per 1 g HPD100 resin; the optimal desorption technology was that the volume fraction of desorption solvent ethanol was 75％; volume flow rate of desorption was 3 BV/h; the volume of desorption solvent was 5 BV. The average desorption retention rate of TSTR was 77. 96％ in technology validation (RSD=0. 46％, n=3) and the purity of prepared TSTR in TSTR dry cream was 52. 47％ (RSD =1. 53％, n=3). CONCLUSIONS: The optimal purification technology is stable, feasible and suitable for the separation and purification of TSTR.

Objective To compare the effects of different extracts of Rheum palmatum on weight and tissue structure of hypothalamusand and pituitary of adult female rats,and screen the main reproductive toxicity extract.Methods The water,chloroform,ethyl acetate,and n-butanol extracts and water-soluble substance of R.palmatum were prepared by polarity gradient extraction method.Female adult rats were randomly divided into blank control group,rhubarb water extract group,and different extracts groups.The dosage of all the groups was equivalent to 4.00 g/kg crude rhubarb.Rats were administered with extracts by gavage for 60 d.Body mass growth rate of rats were calculated before and after administration.The pathological changes of hypothalamic arcuate nucleus neurons and pituitary gonadal cell were observed with light microscope.Results Compared with the blank control group,the body mass growth rate of rhubarb water extract group was decreased (P ＜ 0.05),while those in the different extracts groups were increased (P ＜ 0.01);The hypothalamic arcuate nucleus neurons of rhubarb water extract group showed chromatin marginalization,nissl substance dissolving,fuzzy boundary of nuclear membrane,as well as hell cells,and the total number of adenohypophysis cells reduced and the cells arranged in irregular.However,there were no apparent pathologic changes in different extracts groups.Conclusion Rhubarb water extract administration by long-term dose can reduce weight growth rate and result in pathologic changes of hypothalamic arcuate nucleus and adenohypophysis,while the different extracts can increase weight growth rate significantly and have little effects on the organizational structure ofhypothalamic arcuate nucleus and adenohypophysis.

OBJECTIVE:To investigate the effect of total saponins of Man medicine Thladiantha dubia root(TSTR)on the ex-pressions of CD3+,CD4+and CD8+ in spleen tissue of rats with type Ⅱ collagen-induced arthritis (CIA),and explore its mecha-nism in the treatment of rheumatoid arthritis(RA). METHODS:8 rats were taken as normal control group(NC group),the others 72 rats were injected mixture of bovine type Ⅱ collagen and Freund's complete adjuvant in tail and back to induce CIA model. The 50 modeled rats were randomly divided into model group(MC group),tripterygium polyglycoside(TG)group(12 mg/kg,posi-tive control),TSTR low-dose,medium-dose,high-dose groups (20,40,80 mg/kg),10 in each group. Rats in medicine groups were intragastrically administrated for 35 d,once a day;rats in NC group and MC group were intragastrically administrated equal volume of distilled water. Ankle swelling degree of rats was determined,arthritis indexes were calculated,HE staining was used to observe the lesions in synovial tissue,and immunohistochemistry was used to detect the expressions of CD3+,CD4+,CD8+ in spleen tissue of rats. RESULTS:After 35 d of administration,ankle swelling degree,arthritis indexes,CD4+ expression in spleen tissue,and CD4+/CD8+ ratio in MC group were significantly higher than NC group (P<0.05 or P<0.01),expressions of CD3+, CD8+were significantly lower than NC group(P<0.05);and there was congestion and massive inflammatory cell infiltration in sy-novial tissue. The ankle swelling degree,arthritis indexes,expression of CD4+ in spleen tissue,and CD4+/CD8+ ratio in administra-tion groups were significantly lower than MC group(P<0.05);the expressions of CD3+,CD8+ in spleen tissue in administration groups were significantly higher than MC group (P<0.05);TG group,TSTR medium-dose and high-dose groups showed mild congestion and a small amount of inflammatory cell infiltration in synovial tissue of rats,and TSTR low-dose group showed no ob-vious congestion or inflammatory cell infiltration. CONCLUSIONS:Up-regulating the expressions of CD3+,CD8+ and down-regu-lating the expression of CD4+ may be one of the mechanisms of TSTR in the treatment of RA;and with best efficacy when the TSTR dose of 20 mg/kg.

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

OBJECTIVE:To observe the effects of total organic acid of Thladiantha dubia fruit(TOATF)on coagulation time and hemorheology of rats with cold coagulation and blood stasis. METHODS:60 rats were randomly divided in normal control group (water),model control group (water),aspirin group (positive drug,50 mg/kg) and TOATF low-dose,medium-dose and high-dose groups(50,100,200 mg/kg)with 10 rats in each group. Except for normal control group,cold coagulation and blood stasis model was induced by 4 ℃ water bath and subcutaneous injection of adrenaline hydrochloride,and then given correspon-dence medicine intragastrically,once a day,for consecutive 14 d. 24 h after last administration,blood samples were collected from aorta abdominalis. The coagulating time(CT),erythrocrit(HCT),plasma viscosity(PV),prothrombin time(PT),thrombin time (TT),activated partial thromboplastin time (APTT),platelet aggregation rate (PAR) and whole blood low-shear,middle-shear and high-shear viscosity were measured and blood sedimentation equation K value was calculated. RESULTS:Compared with nor-mal control group,CT,PT,TT and APTT of model control group were shortened,and HCT,PV,blood sedimentation equation K value,PAR and whole blood low-shear,middle-shear and high-shear viscosity increased(P<0.05 or P<0.01). Compared with model control group,CT,PT and APTT prolonged in aspirin group and TOATF medium-dose and high-dose groups,and PAR de-creased;PT of treatment groups prolonged,while HCT,PV,blood sedimentation equation K value and whole blood low-shear and middle-shear viscosity decreased(P<0.05 or P<0.01). CONCLUSIONS:TOATF has obvious improvement effects on anticoagula-tion and hemorheology in rats with cold coagulation and blood stasis.

Objective To investigate the effects of Additive Foshousan(AFSS) on the serum levels of estradiol and progesterone in rats with primary dysmenorrhea. Methods SD rats were divided into six groups, each 10. Group 1 was normal control , groups 2,3 and 4 were treated with low, middle and high dose of AFSS(0.75, 1.5 and 3.0 g/kg) respectively, group 5 as positive controls was treated with Yuanhuzhitong tablets (1 mg/kg)and group 6 served as model control. Except the normal control group, all rats were injected diethylstilbestrol and oxytocin to establish primary dysmenorrhea model. The levels of estradiol(E2)and progesterone(P) in rats serum was determined by ELISA method and the ratio of E2/P was calculated. Results In middle and high dose of AFSS groups, the level of E2(48.27±6.42)pg/L,(47.51±7.03)pg/L respectively were lower than that in model group(54.47±9.12)pg/L and the difference was statistically significant(P<0.05). In low dose of AFSS group, the level of E2 was(50.83±6.26)pg/L and the difference was no statistically significant compared with model group. In all doses of AFSS groups,The content of P(687.41±21.14)ng/L, (720.47±41.03)ng/L, (719.78±32.01)ng/L respectively were higher than that in the model control group (667.32±46.51)ng/L and the difference was statistically significant(P<0.05 or<0.01). In middle and high dose of AFSS groups, the content of P were higher than that of Yuanhu-Zhitong tablets group(699.31±36.31)and the difference was statistically significant(P<0.05). Conclusion To reduce the content of E2, increase of P the content and decrease ratio of E2/P is one of the mechanism for AFSS to treat primary dysmenorrhea.

Objective To investigate the therapeutic effect of Saccharomyces boulardii on antibiotics associated diarrhea (AAD)in infant pneumonia.Methods 302 hospitalized infant patients (1 month-3 years) with pneumonia but without gastroenteric disease were selected , and antibiotics were administrated intravenously at least 5 continuous days for each patient .Patients were all giv-en antibiotics and randomly divided into three groups:control group with no Saccharomyces boulardii administration ( group A, 60 ca-ses);Saccharomyces boulardii were applied as addition on the occurrence of diarrhea (group B, 92 cases), and Saccharomyces boular-dii and antibiotics were co-administrated (group C, 150 cases).Incidences of AAD in all groups were carefully examined and differ-ences of therapeutic effect between groups were compared and analyzed .Results The incidence of AAD in Group C was significantly lower than that in other groups .In term of diarrhea severity, no significant difference was observed in all groups (P>0.05).Howev-er, duration of diarrhea showed significant different between groups (P<0.001):group A with the longest duration and group C with the shortest .The results indicated that the total efficiency of anti-diarrhea in group C was significantly higher than that in control group (P<0.05).However, group B presented no significant difference compared with neither control group nor group C in term of therapeu -tic effect on diarrhea .Conclusion Administration of Saccharomyces boulardii Sachets could shorten the duration of diarrhea on the oc-currence of AAD in infant pneumonia .According to our results , Saccharomyces boulardii was effective in both preventing the develop-ment of AAD and shortening duration of diarrhea , and therefore improved therapeutic effect on ADD .

BACKGROUND: The effective fraction of stem and leaf of Scutellaria baicalensis, total flavones from stem and leaf of Scutellaria baicalensis (SSTF) can improve blood flow of coronary artery and cerebral blood flow,prolong the time of cruor and prothrombin and decrease blood lipid.OBJECTIVE: To observe the effect of SSTF on blood pressure of normal rats and rat models of renal artery stenosis.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Traditional Chinese Medicine, Chengde Medical College (Key Laboratory of Research and Exploiture of Traditional Chinese Medicine in Hebei Province) MATERIALS: Totally 70 Wistar rats, aged 2 months old , of clean degree, with body mass of (250±50) g, male and female in half, with permission number of 01-3003 , were provided by the Experimental Animal Research Institute, Chinese Academy of Medical Sciences. SSTF was provided by the phytochemical laboratory of Institute of Traditional Chinese Medicine, Chengde Medical College; Niuhuangjiangya pian was produced by Shiyitang Pharmaceutical Factory, Hayao Company (batch number:9802304).METHODS: The experiment was conducted in Institute of Traditional Chinese Medicine, Chengde Medical College from January to October 1999. The rats were raised in the environment of (22±1) ℃, with relative humidity of 50%-55%, of daylight and darkness for 12 hours for each. ①In the experiment 1, we studied the effect of SSTF on the blood pressure of normal rats. Totally 30 Wistar rats were randomly divided into SSTF group of 100 mg/kg (n=10, 100 mg/kg SSTF), SSTF group of 50 mg/kg (n=10, 50 mg/kg SSTF), and normal saline control group (n=10, 0.5 mL /100 g normal saline). Before being administrated, rats were anaesthetized by intraperitoneal injection of pentobarbital sodium. Blood pressure was recorded by MecLab/4e four-channel physiologic instrument. Then intragastric administration of SSTF and normal saline were performed on the rats in SSTF groups and normal saline control group respectively. Thirty minutes later, the changes of blood pressure of rats in each group were observed successively. ②In the experiment 2, we studied the effect of SSTF on the blood pressure of model rats with hypertension induced by renal artery stenosis. The rest 40 Wistar rats were randomly divided into SSTF group of high dosage, SSTF group of low dosage, normal saline control group, Niuhuangjiangya pian group, with 10 rats in each group. Rats in each group were created into rat models with hypertension induced by renal artery stenosis according to methods from the literatures. Two weeks after operation, intragastric administration of 100 mg/kg SSTF, 50 mg/kg SSTF, 0.5 mL/100 g normal saline and 1 200 mg/kg Niuhuangjiangya pian were daily performed on the rats in four groups respectively for one month successively. Carotid blood pressure of rats in all groups was measured.MAIN OUTCOME MEASURES: ① The effect of SSTF on the blood pressure of normal rats. ②The effect of SSTF on the blood pressure of model rats with hypertension induced by arteria renalis stenosis.RESULTS: ① Effect of SSTF on the blood pressure of normal rats: Blood pressure of SSTF group of 50 mg/kg and SSTF group of 100 mg/kg presented a little decrease compared with before administration. There was no significant difference when compared with normal saline control group (P ＞ 0.05). ②Effeet of SSTF on the blood pressure of model rats with hypertension induced by arteria renalis stenosis: Blood pressure of rats with hypertension in normal saline control group was significantly increased after modeling; Blood pressure of rats with hypertension in SSTF group of 50 mg/kg , SSTF group of 100 mg/kg and Niuhuangjiangya pian group was significantly lower than that in the normal saline control group, especially significantly in SSTF group of 100 mg/kg [(126.3±14.5),(120.2±15.9),(127.8 ±23.4 ) ,(139.6±15.8) mm Hg; P ＜ 0.05-0.01].CONCLUSION: SSTF has the significant hypotensive effect on model rats with hypertension; it does not have obvious effect on normal rats.

BACKGROUND: Yishen jiangu granule is one of sheer traditional Chinese drugs, which has good effect of tonifying kidney, strengthening bone, reinforcing the spleen, nourishing qi and improving the body condition, mainly used to prevent the lassitude, rickets, osteomalacia and calcium-deficiency induced by kidney-deficiency.OBJECTIVE: To observe the preventive effect of yishen jiangu granule on the reproductive organs, physical strength as well as the ability of stress, and compare with the effect of longmu zhuanggu preparation which was characterized by invigorating spleen and replenishing qi, and tonifying kidney and replenishing essence.DESIGN: Completely randomized controlled design.SETTING: Institute of Traditional Chinese Medicine, Chengde Medical CollegeMATERIALS: A total of 180 Kunming mice, weighing 25-30 g, of half gender, of clean grade, were selected in this study. Longmu zhuanggu granule, which was composed of root of tangshen, tuckahoe, rhizome of lagehead atractylodes, huai yam, astragalus root, schisandra fruit, keel,oyster shell, tortoise plastron, dwarf lilyturf, Chinese date, licorice root,vitamin D, etc., was provided by Wuhan Jianmin Pharmaceutical Factory (batch number: 960618). Yishen jiangu granule, which was composed of 6 kinds of traditional Chinese medicine such as dark plum, Chinese wolfberry, oyster shell, etc., was manufactured by Institute of Traditional Chinese Medicine of Chengde Medical College (batch number: 950510).Hydrocortisone injection was produced by Bohai Pharmaceutical Factory of Tangshan city (batch number: 940303).METHODS: The experiment was completed in Institute of Traditional Chinese Medicine, Chengde Medical College from January to October 1997. ① A total of 180 mice were randomly divided into 6 groups with 30 in each group: normal group (5.0 mL/kg saline, muscle injection); model group (25 mg/kg hydrocortisone for the mice model with kidney-yang deficiency, muscle injection); longmu zhuanggu granule group (5.0 g/kg longmu zhuanggu granule, and 25 mg/kg hydrocortisone); three groups of yishen jiangu granule at different dosages (perfused respectively with yishenjiangu granule at the dosages of 1.25 g/kg, 2.5 g/kg and 5.0 g/kg;meanwhile, 25 mg/kg hydrocortisone, muscle injection). The mice were administered successively for 14 days. ② In 12 hours after giving the last medicine, each group was divided into 3 parts to go on with the following 3 sub-experiments: Firstly, the spermary and uterus of mice were taken out to determine their weight, then calculate the indexes (mass of uterus/body mass of mice); secondly, the mice were put into the water [water depth:30 cm, water temperature: (20±0.5) ℃], burden of tail of mice was weighed 10% of the body mass, then timed immediately, when the mice could not immerge from the water after their heads sank into the water for 10 seconds.It showed that they were exhausted (the time was regarded as the swimming time of mice); thirdly, 10 g sodalime was added into each bottle with wide neck whose volume was 150 mL, then one mouse was put into each of them,covered them and timed immediately. The breath of the mice was observed until they stopped breathing, then timing was over. This was the bearing time of mice on anoxia under normal pressure. ③ Difference of statistical significance between groups was determined by t-test.MAIN OUTCOME MEASURES: The mass indexes of reproductive organs, swimming time and the bearing time on anoxia under normal pressure of the mice in all groups.RESULTS: A total of 180 mice were all involved in result analysis. ① The mass indexes of spermary in mice in normal group, longmu zhuanggu granule group and yishen jiangu groups at high and middle dosages were significantly higher than those in model group (P ＜ 0.05-0.01); the mass indexes of uterus in mice in normal group, longmu zhuanggu granule group and yishenjiangu groups at high dosage were significantly higher than those in model group (P ＜ 0.01). ② The swimming time of the mice in model group was significantly shorter than that in the other 5 groups (P ＜ 0.01). ③The bearing time on anoxia under normal pressure of the mice in model group were significantly shorter than that in the other 5 groups (P ＜ 0.01).CONCLUSION: Yishen jiangu granule has the remarkable effect on kidney-tonifying. It can remarkably relieve the reproductive organ atrophy of the mice with kidney-yang deficiency, has remarkable effect on antifatigue, and also can increase the ability of stress.

BACKGROUND: Qinggong changchun dan (QGCCD) capsule has efficacy in profiting kidney-yang and bettering muscle or bone, it could treat debility, deficiency of energy, forgettery and tiredness and the ache of waist or knee in clinic.OBJECTIVE: To study the effect of QGCCD capsule on stress capability of mice with kidney-yang deficiency.DESIGN: Completely randomized controlled trial.SETTING: Institute of Traditional Chinese Medicine, Chengde Medical College (Key Laboratory of Research and Exploiture of Traditional Chinese Medicine in Hebei Province).MATERIALS: Totally 60 stirp Kunming mice, weighting 19-21 g, of either gender, of grade Ⅱ, were selected in this study.METHODS: The experiment was conducted in the Department of Pharmacology and Toxicology, Institute of Traditional Chinese Medicine of Chengde Medical College from February 2002 to December 2002. Totally 160 Kunming mice of either gender were randomly divided into three groups:Swimming endurance group (n=60), tolerance of hypoxia at normal pressure group (n=50) and single macrophage function group (n=50); and mice in each group were divided into 5 subgroups, including 0.25, 0.5,1.0 g/kg QGCCD groups, model control group and normal control group.Mice were fasted for 12 hours and injected with 25 mg/kg hydrocortisone for 7 successive days to establish yang-deficiency model. Various dosages of QGCCD were given with the same volume of 20 mL/kg for 7 successive days. Mice in model control group were treated with the same volume of saline. Forty minutes later, testes of swimming tolerance, tolerance time of anoxia and engulfment capability of macrophage were performed on mice [To take 20 μL blood in the eyepit vein plexus after the mice were injected 2 minutes and 10 minutes later respectively, and to count the charcoal particle expurgation index (K) of macrophage] to observe effect of various dosages of QGCCD on swimming tolerance, tolerance time of anoxia and engulfment capability of macrophage.RESULTS: Data of totally 160 mice was entered the final analysis without normal control group were longer than those of mice in model control group [(10.94±3.79), (13.68±5.62), (14.58±5.49), (16.12±2.35), (6.45±4.87) minutes;ia of mice in 0.25, 0.5, 1.0 g/kg QGCCD groups and normal control group were longer than those of mice in model control group [(19.45 ±4.63),(21.18±4.25), (22.58±5.21), (23.12±4.78), (12.35±4.89) minutes; t=1.566,macrophage and K index of mice in 0.25, 0.5, 1.0 g/kg QGCCD groups and normal control group were higher than those of mice in model control group (0.023 6±0.010 6, 0.029 1±0.0092, 0.030 8±0.008 6, 0.031 8±0.010 1, 0.012 5±0.008 1; t=2.63, 4.282, 4.898, 4.714, P ＜ 0.05-0.01).CONCLUSION: QGCCD can prolong the swimming time of mice obviously, prolong tolerance time of anoxia according to dosage dependence,increase engulfment capability of macrophags, and strengthen stress capability of mice with kidney-yang deficiency.