BACKGROUND: Since chronic liver disorders are associated with bone loss commonly, it is very significant to probe into the prevention of such bone loss for the treatment of osteoporosis.OBJECTIVE: To study the mechanism of glycyrrhizic acid on prevention and treatment of bone loss induced by liver fibrosis in mice.DESIGN: Randomized and controlled study in which the experimental animals were taken as the objects.SETTING: Experimental animal center, central experimental room and pharmacological research room of a universityMATERIALS: The experiment was performed from January to September 2001. Forty common-grade PCR mice of either sex were employed, weighted varied from 20 to 22 g, provided by Experimental Animal Center of the Institution. According to mass-equivalence principle, 4 groups were randomized, named as control, model group, the treatment with colchicine group(CL group) and the treatment of glycyrrhizic acid group(GA group-) with 10 mice in each group.METHODS: Except the control group, in the model group, CL group and GA group, the subcutaneous injection with 400 g/L carbon tetrachloride prepared with peanut oil was given for 5 weeks to induce liver fibrosis in mice. Afterward, the treatment was applied with carbon-tetrachloride peanut-oil solution 10 mL/kg, colchicum autumnale solution 0. 1 g/kg and glycyrrhizic acid solution 0. 1 mg/kg successively. At the end of the experiment, the eyeball was extirpated for blood collection. The serum was separated to assay the various relevant biochemical indexes of liver injury, observe the changes in liver pathological tissues and measure bone calcium(Ca2+) content of femur and the contents of other bone trace elements as well as bone oxyproline hydroxyproline (Hyp).Effects of glycyrrhizic acid on the contents of bone trace elements in mice of every group.RESULTS: The levels of serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in CL group and GA group were lower than those in the model group( P < 0. 01 ). The level of albumin and ratio between albumin and globulin in the model group were lower than those in CL group and GA group( P < 0.01, P < 0.05) . The content of bone calcium in CL group and GA group were lower than that in the control group ( P < 0. 05),but that was higher than the model group( P < 0.05) . The unit contents of copper(Cu2+ ), magnesium(Mg2+) and zinc(Zn2+) in the right femur of the model group were all higher than the control group ( P < 0.01 ), but the contents of those in GA group were not indicated significant differences compared with the control group ( P > 0.05 ). The liver pathological changes of mice in GA group were obviously milder compared with the model group ( P < 0.01 ) and it was shown with VG staining the severity of hyperplasia of liver collagenous fibers was remarkably milder compared with the model group( P < 0.01).CONCLUSION: The extractive solution of glycyrrhizic acid induces medical metabolic enzyme in the liver, enhances detoxification of liver, protects liver to maintain protein metabolic level and maintains the normal metabolism of bone to promote bone Ca2+ and balance between oxyproline hydroxyproline (Hyp) and trace elements of bone so as to prevent and treat bone loss.

Aim To study the characteristics of osteoporosis induced by long-term administration of prednisone, and investigate the preventive effects of compound stanozolol (CS) on the adverse reactions in male rats. Methods Twenty-four four-month-old male Sprague-Dawley rats were randomly divided into 3 groups with 8 rats per group. Group 1 was control (NS group), other groups were oral gavages prednisone (2.7 mg?kg -1?d -1) first, and then plus vehicle (GC group) or plus CS (combination of Stanozolol 0.5 mg?kg -1?d -1 + Calciofon 0.5 g?kg -1?d -1 + Vitamin D 3 250 U?kg -1?d -1), once a day for 12 weeks. At the experimental endpoint, animals were drawn blood from right ventricle under anesthesia. The tibia, ulna and thighbone were collected to test for parameters related to bone.The undecalcified longitudinal proximal tibia metaphyseal sections were cut and stained with Masson-Goldner's Trichrome (5-?m thickness) for osteoclasts analysis, unstained sections (8-?m thickness) for the cancellous bone histomorphometric and fluorochrome labeling analysis. The measured parameters were used to calculate the percent trabecular area (Tb.Ar%), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular separation (Tb.Sp), percent labeled perimeter (L.Pm%), Mineral apposition rate (MRA), bone formation rate (BFR/TV, BFR/BV, BFR/BS), unit area osteoclast number (Oc.N/BV) and Percent osteoclast number surface perimeter (OcP/BS). The desiccant left ulnar was dissolved in hydrochloric acid to test the content of Ca 2+ and hydroxyproline. Length and width of the thigh bone were tested to observe the instance of bone growing. Results Compared with NS group, bone weight, contents of bone hydroxyproline and Ca 2+decreased remarkably in GC group and the bone histomorphometric parameters also showed bone lose significantly.The effects of CS in prednisone rats showed that CS prevented bone loss effectively.Conclusions Bone loss occured in four-month-old male Sprague-Dawley rats after prednisone-treated for 12 weeks. CS could prevent bone loss effectively by inhibiting bone resorption and advancing bone formation.

AIM To study the relationship between hepatic fibrosis and osteoporosis and to determine the preventive effects of Glycyrrhizin on bone loss in mice. METHOD Forty PCR Mice were randomly divided into 4 groups. Group A mice were controls; Group B mice were given sc injection of 40% CCl 4 10 ml?kg -1 once per 5 days as fibrosis model group; Group C were given orally colchicine of 0 01 mg?kg -1 plus CCl 4 sc, Group D were given Glycyrrhizin(GL) of 100 mg?kg -1 orally plus CCl 4 sc. The four groups were treated for 42 days. The liver injury indexes were measured and the mineral elements and Hydroxyproline of the femur were determined. RESULT Compared with group A, the serum enzymes of alanine aminotransferase (ALT), aspartic acid aminotransferase (AST) were markedly increased and serum albumin (Alb) and A/G(Alb/(total protein Alb) were decreased significantly in group B whose liver slides also showed typical liver cirrhosis. The dried weight of femur in the cirrhosis mice was markedly reduced and the bone Calcium content and bone Hydroxyproline content were also significantly decreased in group B. Bone copper and bone magnesium were increased in group B. In group D, GL inhibited markedly the decrease of the serum enzymes and increased Calcium content and Hydroxyproline content of the bone compared with group B. The bone mass loss was prevented effectively by Glycyrrhizin. CONCLUSION The bone mass was lost in mice with chronic hepatic injury induced by CCl 4 and Glycyrrhizin can prevent bone loss which was accompanied by chronic hepatic injury in mice.

AIM: To study the time dose effects of liver damnification induced by CCl 4 in rats. METHODS: The SD rats were given sc injection 60% CCl 4 twice a week to induced liver damage. After the first week, one proportion animals were killed. The contents of serum alanine aminotransferase (ALT), aspartic acid aminotransferase (ALT), hyaluronic Acid (HA), total protein (TP) and albumin (Alb) were investigated, and the thymus, liver, spleen, kidney, and the adrend gland were weighed. On the other hand, the liver tissue was studied with histopathological to observe the degrees of inflammatory activity and fibrous hyperplasia. In the coming the third, fifth, seventh, ninth week, the different proportion animals were done as the above. RESULTS: In the first week, the level of serum ALT were significantly increased; in the third week, the hepatic cells were occurred fattiness denaturalization; in the fifth week, the histopathological showed the necrotic hepatic cells obviously; in the seventh week, the histopathological displayed that the liver turned into fibrotic; and in the ninth week, the liver changed into hepaticfibrosis induced by CCl 4. CONCLUSION: The injection (sc) of 60% CCl 4 will cause diverse hepatic injury with the dissimilar durative time in rats.

AIM:To study the (rat's) osteoporosis induced by long-term administration of prednisone,and investigate the prophylactic effects of Salvia miltiorrhiza Beg in male rats. METHODS: Twenty-four SD rats were randomly divided into four groups such as control group,prednisone group and Danshen(Salvia miltiorrhiza Bge) group,with 8 rats per group.control group was treated with normal saline,other groups with prednisone at dose of(2.7) mg/kg first,and then prednisone group with normal saline group with Salvia miltiorrhiza Bge at dose of 5.0 g/kg,respectively,once a day for 12 weeks.At the experimental endpoint,bone histomorphometric analysis of thighbone were performed in undecalcified sections,the contet of Ca~(2+) and oxyproline hydroxyproline of ulnar bone were to be measured.length and width of the thighbone were to be tested. RESULTS: Bone histomorphometric parameters showed bone loss was significant to prednisone group(P

AIM: To study the cerebra damaged by long-term administration of prednisone,and investigate the preventive effects of compound danqi (CD) in rats. METHODS: 40 SD rats were randomly divided into 5 groups with 8 rats each group. Group 1 was control (NS group),other groups were oral gavages prednisone ( 2.7 mg?kg -1 ?d -1 ) first,and then plus vehicle (GC group),or plus CD at dose of 2.5 , 5.0 or 10.0 g?kg -1 ?d -1 ,respectively,once a day for 12 weeks. At the experimental endpoint,animals were drawn blood from right ventricle under anesthesia. The left half cerebrum was milled in 10% homogenate to test the content of mono-amine oxidase (MAO),superoxide dismutase (SOD) and acetylcholinesterase (AchE). The right half cerebrum was for histological observation. RESULTS: The concentration of SOD and AchE decreased,yet MAO increased significantly in GC group. Moreover,histopathology showed that the structure of cerebrum cortex became thinner and hippocampi was in disorder. Dropsical and necrotic nerve cells were found. Yet CD could prevent the changes of nerve centre in prednisone tats. CONCLUTION: Cerebra damage occurs in four-month-old male Sprague-Dawley rats after prednisone-treated for 12 weeks. The treatment of CD in different dose can prevent the damage.

BACKGROUND: Chinese herbs of common yam rhizome, balsampear fruit and bagasse fiber have good effects on decreasing blood glucose and lipid, but its mechanism is unclear.OBJECTIVE: To observe the effect of compound preparation of common yam rhizome and balsampear fruit on blood glucose, lipid, blood insulin and anti-infection of rats with type 2 diabetes mellitus (DM).DESIGN: Randomized controlled animal study.SETTING: Technological Developing Center, Pharmacological Department, Experimental Animal Center, and Central Laboratory of Guangdong Medical College.MATERIALS: A total of 80 female SD rats with 4 months old and of SPF grade were selected in this study. Flumamine (Jilin Dongbeiya Pharmaceutical Co., Ltd., batch number: 040126); total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and triacylglycerol (TG) (Beijing Zhongsheng Biotechnological Co. Ltd.); Surestep Life scan blood glucose meter and test paper (Johnson Company, USA); insulin radio-immunity kit (Shanghai Navy Medical Institute); UV-3010 ultraviolet spectrophotometer (Japan Shimadzu); compound preparation of common yam rhizome and balsampear fruit (Technological Developing Center of Guangdong Medical College, batch number: 040321); suspension was extracted from common yam rhizome, balsampear fruit and dietary fiber of bagasse through water with 1 kg/L raw materials.METHODS: Animal breeding and samples collecting were carried out in the Experimental Animal Center and Technological Developing Central Laboratory of Guangdong Medical College from June 2004 to December 2005; meanwhile, detection of marker was carried out in the Pharmacological Department and Central Laboratory of Guangdong Medical College. ①Twenty rats were randomly selected as normal control group and perfused with 5 mL/kg saline every day. Other 60 rats were perfused with 5 mL/kg fat emulsion once a day for 4 weeks, and then, rats were fasted for 12hours and peritoneally injected with 2 g/L streptozotocin (30 mg/kg). Rats in normal control group were peritoneally injected with the same volume of citromalic acid buffer. Three days later, blood glucose was measured ranomly and measured again after 2 weeks. If level of blood glucose was igher than 13.5 mmol/L or urinal glucose was ＞ ++ for two weeks, the models were successful (n=48). ② According to random lot method, 48 rats were divided into three groups: model group, flumamine group and comound preparation group with 16 in each group. Rats in model group were perfused with 5 mL/kg fat emulsion; moreover, rats in flumamine groupr and compound preparation group were perfused with 5 mL/kg fat emulsion and then with 1 mg/kg flumamine and 5 mL/kg compound preparation including 1 kg/L raw drug, respectively. Rats in normal control group were perfused with 5 mL/kg saline. All rats in each group were perfused once a day for 6 weeks in total. ③ Value of blood glucose was measured at one day before the experiment finished. Twelve hours after fasting, level of plasma insulin was measured with radio-immunity method; levels of plasma total protein and albumin were measured with spectrophotography; levels of TG, TC and HDL-C were measured with the related kits. ④ Measurement data were compared with analysis of variance (ANOVA). Levene's test was firstly used to evaluate regularity of variance. Bonferroni test was used for regular variance; however, Tamhane's T2 was used.MAIN OUTCOME MEASURES: Effect of compound preparation on levels of blood glucose, insulin, lipid and plasma protein of rats with type 2 DM.RESULTS: Twelve rats were lost because of failure in modeling, and 4rats in model group and 2 in flumamine group died during the experiment,respectively. Therefore, 62 rats were involved in the final analysis. ①Measurement of fasting blood glucose and plasma insulin: Value of fasting blood glucose in normal control group was lower than that in other three groups (t=2.673-4.224, P ＜ 0.05-0.01), but level of plasma insulin was higher than that in other three groups (t=3.780-5.824, P ＜ 0.05-0.01).Fasting insulin in model group was lower than that in compound prepara tion group (t=2.825, P ＜ 0.05); fasting blood glucose was higher than that in flumamine group and compound preparation group (t=3.906, 3.056, P ＜ 0.05); * level of insulin in flumamine group was lower than that in compound preparation group (t=3.014, P ＜ 0.05); level of fasting blood glucose in flumamine group was close to that in compound preparation group (P ＞ 0.05). ② Measurement of lipid: Levels of TC and TG in normal control group were lower than those in other three groups, but level of HDL-C was higher than that in other three groups (t=2.521-4.892, P ＜ 0.05-0.01).Plasma TC in model group was higher than that in flumamine group and compound preparation group (t=2.466-2.512, P ＜ 0.05), value of TG was higher than that in compound preparation group (t=2.612, P ＜ 0.05), and level of HDL-C was lower than that in compound preparation group (t =3.688, P ＜ 0.05). Plasma TG in flumamine group was higher than that in compound preparation group (t=2.620, P ＜ 0.05). ③ Measurement of plasma protein: Levels of plasma total protein were close to each other (P ＞ 0.05). Plasma albumin in normal control group was higher than that in model group and flumamine group (t=3.773, 3.104, P ＜ 0.05), but that was close to that in compound preparation group (P ＞ 0.05). Ratio between albumin and globulin in normal oln that in other groups (t=2.830-3.056, P ＜ 0.05). Level of plasma albumin and ratio between albumin and globulin were lower in model group than those in compound preparation group (t=2.604, 3.808, P ＜ 0.05).CONCLUSION: Compound preparation can decrease levels of blood glucose and lipid, increase content of insulin, and improve anti-infection ability of rats with type 2 DM.

BACKGROUND: Long-term large dose application of glucocorticoid can cause osseous loss and even femoral head necrosis,which is one of the reasons of pharmaceutical damages. Researches on its intervention have practical significance.OBJECTIVE: To study the osteoporosis induced by long-term large dose administration of glucocorticoid, and investigate the preventive effects of compound danshen(CD) in male rats.DESIGN: A randomized and controlled study by employing experimental animals as subjectsSETTING: An Experimental Animal Center, a Central Laboratory and an Institute of Pharmacology of a Medical CollegePARTICIPANTS: The study was conducted in the Experimental Animal Center, the Central Laboratory and the Institute of Pharmacology of Guangdong Medical College between 2002 and 2003. Totally 40 SD rats were employed.INTERVENTION: SD rats were treated with prednisone(2.7 mg/kg per day) by oral gavages and CD including danshen huangqi , baishu and yinyanghuo at dose of 2.5 g/kg per day,5.0 g/kg per day or 10.0 g/kg per day respectively,once a day for 12 weeks. At the experimental endpoint,the impacts of long term large dose (beyond physiologic dose) application of glucocorticoid on bone metabolism and the preventive effects of CD were observed through the measurement of the static and dynamic indicators for bone growth in un-decalcified superior tibia,the detection of Ca2 + and hydroxyproline contents in ulna,and the length and width of thighbone.MAIN OUTCOME MEASURES: Principal consequences: the impacts of CD on quantitative static and dynamic parameters of osseous morphology in rats with prednisone-induced osteoporosis; Secondary consequences: the comparison of the impacts of CD on bone biochemical indictors and femoral physical indicators in rats with prednisone-induced osteoporosis.RESULTS: In glucocorticoid control group (GC group),bone mass significantly decreased(P<0. 01); as indicated by bone morphological indicators,the number of bone trabecula[(1.98±0.20) / mm]and the percentage of bone trabecular size [(8.83 ±0.98)%] significantly reduced; the ratio of osteogenesis rate at bone surface (8.91±3.97) /neogenesis bone trabacular size to total bone trabecular size(332. 8±142.5)/neogenesis bone trabecular size to bone size(29.6±13.2) significantly decreased; bone absorption perimeter significantly increased(P<0. 01); osseous content in ulna reduced[ (0. 155±0. 01) g]; and femoral length[ (32.64±0.51) mn]significantly shortened (P<0. 05) . But in CD group,CD had certain preventive effects on bone injury induced by prednisone while there was no significant difference among each subgroup with different dose.CONCLUSION: Long-term application of prednisone can significantly inhibit bone growth and induce bone loss. CD has favorablepreventive effects on bone loss through its promotion of osteogenesis and inhibition of osteoclast bone resorption.

AIM: To study the effects of Ginseng fiber on hepatic fibrosis induced by CCl 4 in mice. METHODS: Forty PCR Mice were randomly divided into 4 the control group, the NS group, the colchicine group and the Ginseng fiber groups. Rats in control group were treated by daily oral gavage with vehicle. Rats in other three groups were given SC injection of 40% CCl 4 10 ml?kg -1 and treated by either daily oral gavage with vehicle, or colchicine at 0.1 ml?kg -1, or Ginseng fiber at 10 g?kg -1 for 42 d. The liver injury indexes were measured. RESULTS: Compared with control group, the serum enzymes of alanine aminotransferase (ALT), aspartic acid aminotransferase(AST) were markedly increased but serum albumin (Alb) and A/G were decreased distinctly in CCl 4 group whose liver slides also showed typical liver cirrhosis. Ginseng fiber markedly prevented CCl 4-induced increases in liver weight, serum ALT and TP. Ginseng fiber lightened the hepatic pathological necrosis resulting from CCl 4. The preventive effect of Ginseng fiber was identical to that of colchicine. CONCLUSION: Ginseng fiber can prevent hepatic fibrosis induced by CCl 4 in mice.

AIM To determine the effects of different dose of cyclophosphamide (CP) on the contents of bone calcium and hydroxyproline and the weight of immune organ thymus. METHOD CP at dose of 1 5, 4 5 and 12 5mg?kg -1 were given to the male rats orally everyday for 15 days respectively. At the endpoint the right femoral was dried constantly and weighed (w), the calcium content (Ca.C) was assayed by the method of Atomic Absorption Spectrometry and the femoral hydroxyproline content (Hp.C) were assayed by Colorimetry. The leukocyte, thymus, liver and spleen were tested and weighed separately. RESULTS CP induced inhibiting effect on body weight, leukocyte and thymus in a dose dependent when compared with the vehicle control. Three doses of CP significantly decreased Ca.C and Ca.C/w of femoral ( P0 05). CONCLUSION CP stimulated bone calcium loss, induced shrinkage of thymus. The proper dose at 4 5mg?kg -1 of CP reduced both of Hp.C and Ca.C in bone leading to osteoporosis which is related to the decrease of bone mineral and bone matrix.