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BACKGROUND: Due to glucocorticoid (GG) is used wildly on clinic, incidence rate of steroid-induced osteonecrosis of femoral head is increased.Apoptosis of bone cells plays a key role in steroid-induced osteonecrosis of femoral head during earlier period.OBJECTIVE: To induce models of femoral head necrosis of rabbits by steroid steroid so as to observe the effect of integrated Chinese and western medicine on preventing apoptosis of steroid-induced osteonecrosis of femoral head.DESIGN: Randomized controlled animal study.SETTING: Pharmacological Department and Anatomy Department of Medical College of Shaoguan University, Pathological Department of Yuebei People's Hospital of Guangdong Province.MATERIALS: The experiment was carried out at the Central Laboratory of Medical College of Shaoguan University from April to July 2005. A total of 30 adult New Zealand white rabbits were selected and randomly divided into blank control group, steroid group and preventive medicine group with 10 in each group.Xuesai tongpian, main component of panax notoginseng saponin, was provided by Yuxi Weihe Pharmaceutical Company Limited (batch number: 200410290; 25 mg/pill); zhibituo pian, main component of shanzha, baizhu, hongqu, was provided by Chengdu Di 'ao Jiuhong Pharmaceutical Factory (batch number: 0410063, 350 mg/pill); alendronate, main component of 4-amido-1-hydroxy butylidene-1, was provided by Hainan Mankexing Pharmaceutical Factory (batch number: 040501, 5 mg/pill); dexamethasone sodium phosphate injection was provided by Henan Tiankang Pharmaceutical Company Limited (batch number: 040905, 5 g/L).METHODS: ① One week later, rabbits in steroid group and preventive medicine group were injected with 1 mL dexamethasone sodium phosphate solution twice a week to establish models of steroid-induced osteonecrosis of femoral head. Animals in blank control group did not model and were injected with the same volume of saline at the same time point. ② Rabbits in preventive medicine group were prefused with 25 mg xuesai tong, 350 mg zhibituo and 5 mg alendronate at the same time for 8 weeks. Animals in steroid group and blank control group did not treat with any medicine. Benzylpenicillin was injected into muscle of animals in each group for infection prevention with the dosage of 50 000 u each rabbit twice a week. ③ One week after discontinue, apoptosis of femoral head was detected with in situ end-labeling (ISEL) method. Ten areas were selected from each carry sheet glass to calculate numbers of apoptosis in every 100 cells. Pathological changes of ultrastructure were observed with double staining of uranium acetic acid and lead citromalic acid.MAIN OUTCOME MEASURES: ① Results of apoptosis of femoral head; ② Results of pathological changes of ultrastructure of femoral head under electron microscope.RESULTS: A total of 30 adult New Zealand white rabbits were selected in this study. Because one rabbit in preventive medicine group died during process, 29 rabbits were involved in the final analysis. ① Results of apoptosis of femoral head: Buffy granulosum positive cells, i.e. apoptosis cells,were observed in nucleus in steroid group. Numbers of apoptosis every 100cells were more in steroid group than those in blank control group and preventive medicine group (24.14±5.46, 2.47±0.76, 8.12±3.23, q=8.76, q=13.45, P ＜ 0.01), and numbers in blank control group were similar to those in preventive medicine group (P ＞ 0.05). ② Results of pathological changes of ultrastructure of femoral head under electron microscope: Volume of nucleus in steroid group was decreased; chromatin was aggregated at edge; part of chromatin was dark and thick; density of electron was increased; structure was unclear; characteristics of apoptosis were shown. Nuclear membrane in preventive medicine group was complete under electron microscope, chromatin was steady, and the nucleus was accounted for one third of cell.CONCLUSION: The integrated application of xuesai tong,, zhibituo and alendronate can inhibit hormonal-induced apoptosis of femoral head after necrosis of femoral head induced by steroid steroid.
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BACKGROUND: At present, the researches on steroid-induced avascular necrosis of the femoral head are mostly concentrated on the treatment of formed necrosis. And there are fewer reports on how to prevent the ostoenecrosis of the femoral head in the course of steroid therapy.OBJECTIVE: To observe the effect of preventive medication on femoral head structure and blood flow in steroid-induced osteonecrosis.DESIGN: Randomized controlled experiment.SETTING: Department of Pharmacology, Department of Anatomy, Medical College of Shaoguan University; Department of Nuclear Medicine, Guangdong Province Yuebei People's Hospital.MATERIALS: Thirty adult New Zealand rabbits of either sex, whose body mass was (2.5±0.5) kg.METHODS: The experiment was carried out in the Central Laboratory,Medical College of Shaoguan University, the Department of Electron Microscope, the Northern Campus of Guangzhou, Sun Yat-sen University, and the Department of Nuclear Medicine, Guangdong Province Yuebei People's Hospital from April to July 2005. ①Thirty rabbits were divided randomly into 3 groups with 10 rabbits in each group: control group with intramuscular injection of 1 mL/kg normal saline twice a week and meanwhile, intragastric administration of normal saline(10 mL/d), steroid group with intramuscular injection of dexamethasone sodium phosphate(1 mL/kg) twice a week and meanwhile, intragastric administration of normal saline(10 mL/d),treatment group with intramuscular injection of dexamethasone sodium phosphate(1 mL/kg) twice a week and meanwhile, intragastric administration ofXuesaitong(25 mg/kg), Zhibituo(350 mg/kg) and alendronate(5 mg/kg)daily for 8 weeks. ②After 1 week of drug withdrawal, the blood flow of femoral head was measured in all the rabbits with radioactive microsphere technique, and the histological changes were observed under light microscope and electron microscope.MAIN OUTCOME MEASURES: ①Blood flow of the femoral head in each group.②Histological and morphological changes, and ultrastructure of the femoral head cartilage in each group.RESULTS: ①The blood flow in the treatment group was more than that in the steroid group[(0.261±0.042), (0.197±0.053) mL/(min·g), q=6.10,P ＜ 0.01]. Compared with the control group[(0.243±0.039) mL/(min ·g)],the difference was not significant. ②The number of empty bone lacunae in the treatment group was fewer significantly than that in the steroid group [(15.22±5.49), (24.78±7.87) pieces, q=6.35, P ＜ 0.01]. However, there was no difference between the treatment group and control gruop [(10.38±3.78)pieces].③In the treatment group, the bone cells were normal, the endoplasmic reticula were abundant and the cellular nuclei were of normal shape.In the steroid group, the bone cells contracted in volume, the pyknosis occurred, the chromatin gathered to the edge and the bone lacuna enlarged.CONCLUSION: While using steroid hormone for long, using Xuesaitong,Zhibituo and alendronate may elevate the blood flow of femoral head, improve the tissue structure of bone and prevent or lighten steroid-induced necrosis of femoral head.
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Objective To compare the adherent cells derived from human placenta,umbilical cord blood and bone marrow,and provide laboratory data for clinical application of mesenchymal stem cells.Methods Adherent cells were isolated from human placenta tissues by enzyme digestion,and mononuclear cells(MNC) were isolated from umbilical cord blood(UCB) by 60g/L HES and density gradient centrifugation and MNC were isolated from bone marrow(BM) by density gradient centrifugation,and then these cells were cultured in vitro.Their biological characteristics were studied and compared.Results The adherent cells cultured from human placenta and umbilical cord blood had disparate shape in vitro respectively.And they had some differences in growth and shape from those derived from bone marrow.The adherent cells derived from the three tissues all expressed CD106 and CD44 in immunohistochemistry staining.Conclusion The adherent cells derived from human placenta and umbilical cord blood have the basic features of mesenchymal stem cells.
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The phenotype frequencies of Gc and Tf in blood and bloodstains from the Hannationality in Bejing was studied using the agarose gel simultaneous electrophresisfollowed by the immunofixation method.Their gene frequencies were calculated.
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Objective To understand the effects of various decoration materials and furniture on indoors air quality. Methods Formaldehyde, benzene and ammonia were monitored in indoor air of some residential buildings in Guangzhou. The residential rooms were divided into two groups, one was simply decorated group, the other was completely decorated group and the two groups were compared. The questionnaire was also carried out. Results The average concentrations of formaldehyde of the two groups were 0.338 mg/m3 and 0.027 mg/m3 respectively, the rates of exceeded standard were 57.14%, 6.67% respectively, and the difference between the two groups was obvious (P
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A region of the D17S30(pYNZ22) locus was amplified in DNA from 120 unrelated chinese individuals was carried out. The amplified products were analysed by mini-polyacrylamide gel electrophoresis followed by silver stain. 11 alleles, ranging from 170bp to 870bp in size and from 0. 4 % to 30. 4 % in frequency, was detected. The heterozygosity was 73%, Dp value was 0.938. The family study showed that the Amp-FLP in pYNZ22 locus is inherited according to the Mendelian law. The correct genotyping results can be obtained from very little amount of biological material, such as mixed stains, blood stains, single hair and saliva.
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The polymorphisms of minisatellite p33.6 (D1S111) locus were typed rapidly and accurately us- ing polymerase chain reaction (PCR). The amplified fragments were analyzed by mini- polyacrylamide gel electrophoresis followed by silver stain. The distribution of allele frequencies among 100 unrelated Chinese is reported. 14 alleles, containing 9 to 22 repeat units, had been detected with a heterozygosity of 76%. The allele frenquencies were from 0.5 %to 35.5 %. The size of amplified fragments ranged from 435 bp to 925bp. The discrimination power of p33.6 locus was 0. 916. The Amp --FLP was inherited according to the Mendelian Law. The results showed that the polymorphisms of p33.6 locus can be used for individual identification and paternity test.
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DNA extracted from 100 unrelated Chinese were detected by using the Polymerase Chain Reaction (PCR), mini polyacryl gel vertical electrophorese and silver staining techniques at the p33. 4 locus. Among 100 unrelated individuals, 8 alleles were detected,Foster-Freeman BIOTRAC system indicated that the number of copies of the tandem repeats in different allele was 7, 10 to 15 respectively.There was a rare allele whose copy number was more than 13 but less than 14. The allele fragmehtlength varied from 603 to 1115bp,the allele freqency 0.5 % ~ 53. 5 %,heterozygosity 64 %,DP value84. 5 %. Pedigree analysis indicated that alleles of p33. 4 locus obey Mendel's Laws. Successful typingof various tissues and single hair folicle had confirmed that this technique is suitable for personal identification. In addition, by using Chelex we had established an alterntive method for extracting DNAfrom biological materials,whlch is rapid and easy to perform.