2.Effects of donkey-hide glue reinforcing bone oral solution medicated serum on osteoprotegerin and osteoprotegerin ligand mRNA expression of osteoblast in fetal rats
Lin SHEN ; Jialin WU ; Yuanjun XIA ; Lei LI ; Lan GAO ; Jing XIE ; Piqi ZHOU ; Yanping YANG
Chinese Journal of Tissue Engineering Research 2005;9(42):146-148
BACKGROUND:The therapeutic effects of donkey-hide glue reinforcing bone oral solution on osteoporosis have been determined, but the exact effective mechanism is to be approached. OBJECTIVE: To investigate the effects of donkey-hide glue reinforcing bone oral solution (DGRBOS) medicated serum on osteoprotegerin (OPG)and its ligand(OPGL)mRNAexpression of osteoblast in fetal rats and explore the molecular mechanism of treating osteoporosis with DGRBOS. DESIGN: A randomized controlled trial. MATERIALS: The experiment was carried out from June 2003 to October 2004 in Bone Metabolic Laboratory of Department of Integrative Chinese and Western Medicine, Affiliated Hospital of Tongji Medical College,Huazhong University of Technology and Science. Totally 30 3-month-oldWistar rats (15 males and 15 females) were randomly divided into 3 groups, I.e. DGRBOS group, estrogen group and control group, with 10 rats in every group. 12 clean newborn SD rats were selected to isolate and cul ture osteoblast. METHODS: ①After intragastric administration for 7 days, medicated serum was prepared respectively from the three groups. ②Skull osteoblast isolated from newborn SD rats was made into single cell suspension, then after digestion and passage, the subcultured osteoblast cell was made into cell suspension. The cultured osteoblasts were divided into 5 groups and given equal volumes of drug liquor. The DGRBOS group was given DGRBOS-medicated serum at the concentration of 100, 500 and 1 000 g/L which was diluted by nutrient solution; the estrogen group was given tibolone-medicated serum of 100 and 1 000 g/L; the control group was givenonly culture fluid. Meanwhile every group was given calf serum (100 g/L) for further culture. ③The osteoblast proliferation was measured by antigenic MTT colorimetric analysis and 3H-TdR penetration method. The in tra-cellular BGP contents were evaluated by radioimmunity .The mRNA expression of OPG and RANKL in osteoblast was analyzed by Rt-PCR. ④ One-way analysis of variance was applied to compare data among groups. MAIN OUTCOME MEASURES: mRNA expression of OPG and PAN KL in osteoblasts from fetal rats after intervention by medicated serum ofDGRBOS or Livial. RESULTS: ①The osteoblast proliferation measured by antigenic MTT colorimetric analysis and 3H-TdR penetration method showed that the proliferation in the DGRBOS group and tibolone group was enhanced moresignificantly than that in the control group (P < 0.05-0.01), and reached maximal effect at the concentration of 500 g/L (P < 0.01), but when the concentration was over 500 g/L, the effect tended to saturate. The medicated serum with all concentrations from DGRBOS and estrogen groups could increase the contents of BGP in osteoblasts (P < 0.05). ②The mRNA expression of OPG reached the peak when the DGRBOS medicated serum was 1 000 g/L, and was obviously higher than that at the concentration of 100 and 500 g/L (P < 0.05). The expression in DGRBOS group at the concentration of 1 000 g/L and in the estrogen group at the concentration of 100 and 1 000 g/L was apparently higher than that of the control group (P < 0.01). ③The mRNAexpression of RANKL was the highest in DGR BOS group with 1 000 g/L concentration, and was markedly lower than that of the concentration of 100 and 500 g/L (P < 0.05). The expression in DGRBOS group at the concentration of 1 000 g/L and in the estrogen group at the concentration of 100 and 1 000 g/L was noticeably lower than that in the control group (P < 0.01).CONCLUSION: ①The DGRBOS could remarkably enhance osteoblast proliferation in dose-dependent and a dose-saturable manner, and the effect was close to that of tibolone. ②Partial mechanism of DGRBOS in treating osteoporosis might be promoting osteoblast proliferation and regulating OPG/RANKL expression.
3.Safety, tolerability and pharmacokinetic study of recombinant human parathyroid hormone rhPTH (1-84) in Chinese healthy volunteers.
Qian, LI ; Jian, QIAO ; Jungang, DENG ; Tianshu, ZENG ; Piqi, ZHOU ; Weiyong, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):431-4
The current study was designed to determine the safety, tolerability and pharmacokinetic parameters of recombinant human parathyroid hormone [rhPTH (1-84)] used for the treatment of osteoporosis. In the single-dose format pharmacokinetic study, thirty-six healthy male volunteers received three dose levels of rhPTH (1-84) subcutaneously: 1, 2, and 4 mug/kg. The blood was timing drawn and the serum concentration of rhPTH (1-84) was determined by enzyme linked immunosorbent assay (ELISA). Serum concentration-time curves of PTH (1-84) exhibited a double-peak pattern, the first peak appearing about 10 to 30 min after administration and the second peak occurring about 1.5 to2 h after administration. Serum terminal half-time of PTH (1-84) was approximately 2 h. The parameters indicated the serum levels were directly proportional to the administered dose, with the mean C(max) and AUC(0-24) ranging from approximately 543.47 to 1845 pg/mL and 2358.6 to 9232.12 pg.h.mL(-1) over the dose range. The drug was well tolerated, the clinical symptoms were generally mild and of short duration.
4.Effects of epimedium total flavonoids on bone mineral density and bone metabolism-related indices in primary osteoporosis
Zhexing SHOU ; Lin SHEN ; Yanping YANG ; Jing XIE ; Piqi ZHOU ; Lan GAO
Chinese Journal of Tissue Engineering Research 2009;13(11):2191-2195
BACKGROUND: Previous animal experiments have shown that epimedium total flavonoids can exhibit preventive effects on estrogen-related bone loss, but few data are available in clinical research.OBJECTIVE: To investigate the effects of pimedium total flavonoids on bone mineral density (BMD) inprimary osteoporosis.DESIGN, TIME AND SETTING: A randomized, double-blinded, positively controlled clinical observation was performed at the Department of Traditional Chinese Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between June 2005 and September 2007.PARTICIPANTS: A total of 64 patients with primary osteoporosis consisting of 11 males and 53 females were included in this study. METHODS: All patients were randomly and evenly assigned into a treatment group and a control group. The treatment group was orally administered epimedium total flavonoids, 0.45 g once, three times daily, for a total of 6 months. Simultaneously, the control group was orally administered Gushukang, 10 g once, twice daily, for a total of 6 months. MAIN OUTCOME MEASURES: BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, greater trochanter, and left hip was measured, and simultaneously serum levels of calcium, phosphorus, and alkaline phosphatase (ALP) were detected pdor to and after treatment in each group.RESULTS: All 64 patients with primary osteoporosis were included In the final analysis, The BMD in the lumbar vertebrae (L1-4) was significantly higher in the treatment group than in the control group (P<0.05) and there was no significant difference In BMD in other sites between the two groups (P>0.05). Compared with pdor to drug application, BMD in the treatment group did not present obvious change after epimedium total flavonoids application, while in the control group, BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, and greater trochanter was significantly decreased (P<0.01-0.05). After drug application, serum level of calcium was significantly increased in each group, compared with prior to drug application (P<0.05), and no significant difference existed between the treatment and control groups (P>0.05). Tn the treatment group, serum levels of phosphorus and ALP did not alter significantly compared with prior to epimedium total flavonoids application (P>0.05).CONCLUSION: Epimedium total flavonoids exhibit effective effects on the maintenance of BMD in primary osteoporosis.
5.Actarit vs methotrexate in treating rheumatoid arthritis
Yanping YANG ; Lin SHEN ; Jing XIE ; Piqi ZHOU ; Baoping IU ; Lan GAO
Chinese Journal of New Drugs and Clinical Remedies 2001;20(3):177-179
AIM: To campare the therapeutic effects and security between actarit (MS 932) and methotrexate(MTX) in the treatment of rheumatoid arthritis(RA). METHODS: One hundred and twenty patients with RA were randomly divided into two groups. Eighty patients of actarit group receved actarit, 100 mg, po, tid for 12 wk. The other forty patients received methotrexate 10 mg, po, qw for 12 wk as control. RESULTS: The total effective rates were 73 % for actarit and 78 % for MTX (P>0.05). The adverse reactions were 9 % and 18 %(P<0.05). CONCLUSION: Actarit is as same as methotrexate in the therapeutic effect, but actarit is better than MTX in security.
6.Effect of Xiaoyu Zhixue Tablets on Expression of Membrane Glucoprotein in the Patient of Hemorrhagic Thrombopathy
Lin SHEN ; Di SHEN ; Lan GAO ; Zhongping LIU ; Yanping YANG ; Jing XIE ; Piqi ZHOU
Journal of Traditional Chinese Medicine 1993;0(04):-
Objective:To observe effect of Xiaoyu Zhixue Tablets on expression of platelet membrane glucoprotein in the patient of hemorrhagic thrombopathy and probe into the mechanism of the therapy.Methods:148 cases of hemorrhagic thrombopathy were randomly divided into a Chinese drug group(n=98)treated by Xiaoyu Zhixue Tablets,and a Western medicine group(n=50) treated by adrenosem,vitamine C,K,P.They were treated for 6 months.After treatment the therapeutic effect of hemostasis and the recovery rate of platelet aggregation in the two group were observed and analyzed.Before and after treatment expressions of platelet membrane glucoproteins GP Ⅰ b/Ⅸ,GP Ⅱ b/Ⅲa,GP Ⅰ b,GP Ⅱ b,GP Ⅲ a and p-selectin expression were detected with flow cytometry in the two groups and in 34 healthy persons(normal group).Results:The total effecive rate of hemostasis was 89. 8% in the Chinese drug group and 54.0% in the Western medicine group,and the recovery rate of platelet aggegation was 72.4% in the Chinese drug group and 4.0% in the Western medicine group,with significant differences(both P0.05).Conclusion:Xiaoyu Zhixue Tablets can up-regulate platelet membrane glucoproteins GP Ⅰ b/Ⅸ,GP Ⅱ b/Ⅲ a,GP Ⅰ b,GP Ⅲ a and p-selectin expression,which is possibly one of the mechanisms for treatment of hemorrhagic thrombopathy.
7.Interventional effects of donkey-hide glue reinforcing bone oral solution on the expression of growth factors during fracture healing
Heng LI ; Lin SHEN ; Liqin LI ; Yanping YANG ; Jing XIE ; Piqi ZHOU ; Yi DAI ; Furong LU ; Jialin WU
Chinese Journal of Tissue Engineering Research 2007;11(23):4634-4637
BACKGROUND:Donkey-hide glue reinforcing bone oral solution (DGRBOS) is effective on preventing and treating fracture, but the mechanism of harmacology is still not clear. Vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2) are important cytokines, which can promote blood vessel growth and osseous anabolism during fracture healing.OBJECTIVE: To investigate the effects of DGRBOS on expression of VEGF and FGF-2 in the process of fracture healing of SD rats' fracture of tibia in bony callus, and explore the mechanism of DGRBOS in the treatment of fracture.DESIGN: A completely randomized controlled study.SETTING: Department of Traumatic Orthopedics, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: Ninety female Sprague-Dawley (SD) rats, with a mean weight of (368±40) g, aged about 3 months, wereprovided by Center of Animal Experiment, Tongji Medical College, Huazhong University of Science and Technology.Experimental drug: DGRBOS was donated by Xinjiang Huashidan Pharmaceutical Co., Ltd. The qili linking bone pill,positive control drug, was purchased from Hunan Pharmaceutical Co., Ltd.METHODS: This experiment was carried out in the Laboratory for Bone Metabolism of Integration of Chinese and Western Medicine (Laboratory of Provincial Level), Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from June to November 2005. Transverse midshaft fractures were produced on the right tibia in these rats using three-point bending technique, and 90 rats were randomly divided into three experimental groups:the DGRBOS group (n =30): according to medicine conversional method between human being and animal, 2 mL DGRBOS was fed to every mouse by intragastric administration at 2 vices/day; the qili linking bone pill group (positive control group, n =30): the pills dissolved in distilled water at 225 g/L, and consequently were administered intragastrically into mice at 2 mL/vice and 2 vices/day; normal saline group (negative control group, n =30): normal saline was administered intragastrically into mice at the coordinative capacity and same frequences. Selecting 4, 7, 14,21 and 28 days during the experiment, the immunohistochemistry method was adopted to detect the change of expression of VEGF and FGF-2 through computing value of mean optical degree (MOD) and number of the positive cells.
8.Safety, Tolerability and Pharmacokinetic Study of Recombinant Human Parathyroid Hormone rhPTH (1-84) in Chinese Healthy Volunteers
LI QIAN ; QIAO JIAN ; DENG JUNGANG ; ZENG TIANSHU ; ZHOU PIQI ; LI WEIYONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):431-434
h. The parameters indicated the serum levels were directly proportional to the administered dose, with the mean Cmax and AUC0-24 ranging from approximately 543.47 to 1845 pg/mL and 2358.6 to 9232.12 mild and of short duration.