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Objectives:To find out factors that influence the public's organ donation willingness,and explore the effect on organ donation of soft culture system.Methods:We conducted a survey towards people from typical cities and counties of Zhejiang Province by using self-made questionnaire.We adopt some data processing methods such like entropy weight method.Results:Factors like education,age,traditional concept all can affect the degree of organ donation willingness.Soft culture system has favorable social effects.It is directive in reality.Conclusions:In the humanistic perspective,the construction of organ-donation soft culture system plays an obvious role in improving the degree of social organ donation willingness.It includes education training,humanitarian assistance model radiation,and incentive method.
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Objective To investigate the effects of alcohol extract of Kunmu decoction on proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). Methods Synovial tissues were obtained from patients with active RA received joint replacement or arthroscopy. The surface antigen and the amount of apoptotic cells were determined by flow cytometry. The inhibitive effect was detected by MTT assay. Results The CD90+surface antigen of synoviocytes was (94.78 ± 0.98)%. The inhibitive effect on the proliferation in all treatment groups were in a time-and dose-dependent manner. The apoptosis rate was increased in a dose-dependent manner among all dosage alcohol extract groups. Conclusion Kunmu decoction might inhibit proliferation and induce apoptosis of RA-FLS.
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OBJECTIVE: To study the effects of Tongbi Mixture 2, a compound traditional Chinese herbal medicine for treating rheumatoid arthritis (RA), on immunoregulation of T lymphocytes in rats with collagen-induced arthritis (CIA). METHODS: Forty Wistar rats were randomly divided into normal control group, untreated group, Tongbi Mixture 2-treated group, methotrexate (MTX)-treated group and Tripterygium wilfordii polyglycoside (TWP)-treated group. Except for the rats of the normal control group (injection with normal saline), rats of the other four groups were subcutaneouly multipoint-injected with collagen protein II to induce CIA. The rats were treated with normal saline, Tongbi Mixture 2, MTX tablets and TWP tablets respectively for 36 days. The expressions of CD28 and CD152 were detected by flow cytometry, while the content of tumor necrosis factor-alpha (TNF-alpha) was analyzed by enzyme-linked immunosorbent assay. RESULTS: The expression of CD28 among the three drug treated groups had no statistical difference (P>0.05), while significantly higher than that of the normal control group (P0.05). There were no statistical differences in content of TNF-alpha between the drug treated groups and the normal control group (P>0.05), but the content of TNF-alpha of the drug treated groups was lower than that of the untreated group (P<0.05 or P<0.01). CONCLUSION: Tongbi Mixture 2 can inhibit T lymphocytes through down-regulating the expressions of CD28 and CD152 and the content of TNF-alpha, which may be the major mechanisms in treating RA.
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BACKGROUND: Chinese medicine is rich in recognition and treatment for rheumatoid arthritis(RA) and has achieved attractable progression in researches of recent several years,characterized by variable therapeutic methods, definite therapeutic effects and few toxic side effects. Therefore, it is important scientifically and socially to further search for the effective drugs and methods for the treatment of RA in the field of Chinese medicine. There are few reports on whether Chinese herbs inhibit or block the destruction of cartilage and bone and promote their metabolism and repair for damage or not by variable approaches,targets and links in RA occurrence.OBJECTIVE: To probe into the mechanism of Chinese herbal tongbiling on cartilage destruction.DESIGN: Complete random design and control experimental study based on the experimental animals.SETTING: Golden Chamber teaching and research room of guangzhou university of traditional Chinese medicine.MATERIALS: The experiment was performed in Comprehensive Experimental Room of First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from August 2002 to December 2003,in which,35female Wistar rats were employed,clean grade,weighted(110±20) g,provided by Animal Experimental Room of First Military Medical University of Chinese PLA, with qualified animal No. 2002A041.After 3 days adaptive breeding,Wistar rats were randomized into normal group,model group and total alkaloidal of tongbiling group(TBL group). TBL was extracted in Phytochemistry Room of Tropical Disease Institute of Guangzhou University of Traditional Chinese Medicine.METHODS: The animal model was prepared on rats with collagen-induced arthritis. After treated with gastric infusion in groups,the pathological sections were done on phalangeal joint of rat and chondropathy was observed after routine HE staining. The chondrocytes were cultured. The chondrocytes were stimulated with rrIL-1β in vitro and resulted in matrix decomposition of chondrycyte to prepare pharmaceutical model,TBL of various concentrations added and dexamethasone(DXM) was applied in the control. After 48 hours culture,the upper clear solution if cell was collected and glycosaminoglycan (GAG) content was measured with alcian blue method,and nitric oxygen (NO) content was assayed with method of aqua fortis reduction.MAIN OUTCOME MEASURES:①Primary results: Effect of TBL on phalangeal chondropathy in rats with collagen-induced arthritis and on matrix decomposition of chondrocyte induced by rrIL-1β.②Secondary results: Effect of TBL on induction of NO synthesis from chondrocytes by rrIL-1β.RESULTS:①It was indicated in evaluation on HE pathological section of phalangeal joint: The severity of cartilage destruction in TBL group was less than that in model group(P<0.05).②TBL of various concentrations and DXM inhibited remarkably the decomposition of choncrocyte proteoglycan (P<0.01) and NO synthesis (P<0.01).CONCLUSION: TBL inhibits phalangeal cartilage destruction of rats with collagen-induced arthritis. It is indicated in the results of experiment in vitro that TBL is against the decomposition of choncrocyte proteoglycan,which is probably achieved by inhibiting NO synthesis.
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BACKGROUND: Researches on the pathogenesis and pathological changes of rheumatoid arthritis(RA) have achieved significant progress in recent years. But traditional Chinese medicine(TCM) has unique advantage in RA therapy.OBJECTIVE: To study effects of overall alkali in tongbiling(TBL) on the proliferation of lymphocytes and the transferrin receptor of T lymphocytes (CD71) to explore the mechanism of TBL on the modulation of cell immunity.DESIGN: A completely randomized grouping design and an explorative study by employing cells as subjects.SETTING: Sixth internal medicine department of a TCM university,center of tissue transplantation and immunology college of life science of a university PARTICIPANTS: The study was conducted in the central laboratory (tertiary laboratory of National TCM Administrator) of the first affiliated hospital of Guangzhou TCM medical university between July 2002 and August 2003. Ten clean male SD rats were selected.METHODS: Lymphocyte was separated from rat inguinal lymph node for culture. Concanavalin(ConA) was used for 72-hour stimulation. The impacts of overall alkali TBL on lymphocyte proliferation were tested by MTT.The expression of T lymphocyte CD71 was tested by flow cytometer after 48-hour stimulation of phorbol 12,13 -dibutyrate(PDB) or ConA.MAIN OUTCOME MEASURES: The impacts of overall alkali TBL on lymphocyte proliferation and T cell activation.RESULTS: Different concentration of overall alkali TBL could significantly inhibit the proliferation of lymphocytes under ConA stimulation. PDB and ConA-activated T lymphocyte CD71 + expressions were significantly higher than that of blank control group(P<0.01) . CD3+ CD71 + expressions [(62.03±1.51) %,(25.28±1.57) %,(20. 29±1.72)%] activated by ConA under different concentration of overall alkali TBL(50,100,200 mg/L)were significantly lower than(72.03±1.28)% of BPS-positive control group (P<0. 05). CD3 + CD71 + expressions activated by PDB under 100 mg/Land 200 mg/L of overall alkali TBL were significantly lower than that of phosphate buffer solution (PBS-)positive control group(P<0.05). Different concentration of overall alkali TBL had significant down-regulated effects on CD71 expression in T lymphocyte activated by PDB or ConA and there was also a significant dose-effect relationship(P<0. 05). The inhibition on ConA-activated CD71 expression was stronger than that of PDB.CONCLUSION: Overall alkali TBL can inhibit the abnormal proliferation of T lymphocyte and its mechanism might be realized through its inhibition on transferrin receptor.
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BACKGROUND: Tongbiling is the modified guizhi shaoyao zhimu tang (Decoction of Cinnamon Twig, Peony and Anemarrhena) recorded in Synopsis of Prescriptions of the Golden Chamber. It has been used for rheumatoid arthritis(RA) for more than ten years. Here is the experimental study on effects of tongbiling on interleukin 2(IL-2) and its α-chain, IL-2Rα(CD25), to explain the mechanism of tongbiling in regulating the cellular immunity and resisting RA From the aspects of in vivo and in vitro.OBJECTIVE: To study the effect of tongbiling on IL-2 in collagen introduced arthritis of rats and the influence of total base of tongbiling on expression of IL2-Rα (CD25).DESIGN: A totally randomized and controlled experimental study based on the experimental animalsSETTING: Department of Jinkui of a university and the center of organ transplantation and immunity of another university.MATERIALS: The experiment was carried out from April to July 2001 in the Central Laboratory of the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine. The 36 Wistar rats, clean degree, male,bought from the Animal Experimental Center of Medical College of Sun Yat-sen University, certificate No. 2000A050, weighted(110 ± 20) g, were divided into 6 groups with 6 in each: normal group, model group, methotrexate (MTX) high dosage group, MTX low dosage group, tongbiling high dosage group, and tongbiling small dosage group.METHODS: The experiment in vivo was done with the modeled rats of collagen-induced arthritis. The foot swelling was measured with volume method, the cells of primary synovium cultured, and the upper clear fluid of culture fluid tested with radioimmunoassay to observe the effect of tongbiling on synovial IL-2. The experiment in vitro was done in the way of separating the lymphocytes, giving extraneous stimulation with phorbol 12, 13-dibutyrate (PDB) or concanavalin A(ConA), using bicolor immunofluorescence labeling, testing with flow cytometer to observe the influence of total base of tongbiling on the expression of CD3 +, CD25 +.MAIN OUTCOME MEASURES: The influence of tongbiling on the foot swelling and synovial IL-2 of rats with collagen-induced arthritis.RESULTS: In the experiment in vivo, the high dosage of tongbiling could relieve obviously the foot swelling, compared with MTX, there was no remarkable difference ( P ＞ 0.05) . Both high and low dosage of tongbiling could inhibit the synthesis of synovial IL-2 ( P ＜ 0.01 ). In the experiment in vitro, the total base of tongbiling could extremely inhibit the expression of CD3 +, CD25 + stimulated by ConA, but not effective for that stimulated by PDB plus ionomycin.CONCLUSION: Tongbiling can inhibit the IL-2 synthesis and the signal transduction of IL-2Rα (CD25) activated by ConA, relieving the inflammation of local joint. It provides an experimental evidence for the application of tongbiling in the treatment of RA.
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Objective: The study was to reveal the inhibition of neovascularization by Tongbiling. Methods: Rat model were established by injecting CFA at the root of the tail and divided into model control group, Indomethacin group and Tongbiling group at random. The serum and paws were collected after seven days treatment. 10ng/L VEGF solution was added I into CAM to stimulate the neovascularization, and to observe the influence of each group of serum on the first and second order new vessels. RT-PCR method was taken to determine the expression of mRNA of VEGF and VEGFR-1 of paws in different groups. Results: The number of vessels in the model control group was significantly more than that in the Indomethacin group and the Tongbiling group (P
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[ Objective ] To establish a method for the determination of serum strychnine concentration in rheumatoid arthritis patients treated with Tongbiling prescription and to monitor the serum strychnine concentration. [ Methods ] Serum strychnine was extracted with solid phase extraction column. Reversed-phase HPLC conditions were: Hypersil Division ODS C18 column (150 mm ?4.6 mm, 5?m), mobile phrase being methanol: water (70 : 30) with 0.6 g potassium dihydrogen phosphate and 1.0 g dodecyl sodium sulfonate added in 1000 mL mixture (adjust pH to 4.5 with acetic acid), detecting wavelength at 254nm and a flow rate of 1.0 mL/min at room temperature. [Results] Strychnine has a good linearity in the range of 0.031- 2.000?g?mL-1. The minimum detecting concentration was 0.01?g?mL-1 and the average recovery was over 90%. Intra-day RSD and inter-day RSD ranged from 1.53% to 5.32%. [Conclusion] This method is simple and efficient, and can be used for the determination of strychnine.
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[Objective]To observe the effects of total alkaloids (TA) of Tongbiling (TBL) prescription on proliferation of T lymphocytes and transferring receptor of T lymphocytes (CD71) in rats and to explore the mechanism of TBL in regulating cellular immunity. [Methods]Isolated lymphocytes from mesenteric lymph node were cultured with various concentrations of TBL TA and canavaline A (ConA) for 72 hours; the proliferation of T lymphocytes was observed by MTT method and CD71 expression rate of T lymphocyte activated by phorbol 12,13-dibutyrate (PDB) or ConA for 48 hours was detected by flow cytometry.[Results]Various concentrations of TBL TA inhibited the proliferation of T lymphocytes activated by ConA and decreased CD71 expression rate of T lymphocyte activated by ConA or PDB in a dose-effect manner. Inhibitory effect of TBL TA on CD71 expression rate of T lymphocyte activated by ConA was superior to that activated by PDB. [Conclusion]TBL TA can inhibit the abnormal proliferation of T lymphocytes and its mechanism may be related to the expression of CD71.
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[ObjectivejTo explore the therapeutic mechanism of Tongbiling (TBL) prescription for rheumatoid arthritis (RA) by observing the effects of TBL on inflammation and expression of vascular endothelial growth factor (VEGF) in synovium of rats with collagen-induced arthritis (CIA) .[Methods]CIA rat models were established by subcutaneous injection of type II collagen. Pathological sections of synovium in knee joint were made after medication for 37 days and then stained by HE method and immunohistochemical method respectively. Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in HE-stained pathological sections were observed under microscope and expression of VEGF was observed by semi-quantitative immunohistochemical method. [ Results ] Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in high-dose TBL group were inferior to those in model group (P 0.05); both TBL and Radix Tripterygii Wilfordii at high dose decreased the expression of VEGF (P
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[Objective] To observe the effects of Tongbiling (TBL) prescription in counteracting cartilage destruction in rats with rheumatoid arthritis (RA). [Methods] Collagen-induced arthritis (CIA) rat models were established to observe the therapeutic effects of TBL. After a short-term treatment, expressions of tumor necrosis factor (TNF - a) and transforming growth factor (TGF- P) in cartilage tissue of model group, methotrenxate (MTX) tablets group and Radix Tripterygii Wilfordii (RTW) preparation group were examined by immunohistochemical semi-quantitative assessment method. [ Results ] The number of chondrocytes with positive TNF - a expression was lower and the chondrocytes were stained lighter in normal group than those in model group and low-dose MTX group ( P
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[Objective] To investigate the effect of Erteng Mixture (EM) on activated T-cell CD69 expression and Jurkat cell cycle in rheumatoid arthritis (RA) and to explore its possible cellular immunoregulation mechanism. [Methods] Jurkat cells were cultured with different kinds of components extracted from EM. After 72 hours, proliferation rate of Jurkat cell were detected with thiazolyl blue colorimetry (MTT) to screen the active component. Whole blood from RA patients were incubated in RPMI-1640 culture with various concentrations of chlorofonn component extracted from EM or with Genistein. Thirty minutes later, phytahematoagglutinin (PHA) were added successively into the culture. After 24 hours, CD69 expression rate of T lymphocytes activated by PHA was analyzed by flow cytometry. Jurkat cells were cultured with various concentrations of chloroform component extracted from EM and with methotrexate ( MIX) for 48 hours. After then, Jurkat cell cycle was analyzed by flow cytometry. [ Results ] Chloroform component extracted from EM had the strongest inhibitive effect on proliferation of Jurkat cells. Chloroform component (10 mg/L and 20 mg/L) could markedly inhibit CD69 expression on PHA-activated T lymphocytes. Chloroform component 10 mg/L blocked Jurkat cell cycle at G1 phase, which differed from MTX blocking Jurkat cell cycle at S phase. [Conclusion] Chloroform component extracted from EM could significantly inhibit the early activation of CD4 T lymphocytes in RA and inhibit the energy and material synthesis in T-cell, thus hindering DNA synthesis and decreasing the cellular G2 phase transformation. This study will provide an experimental basis for clinical application of EM in treating RA.
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Objective To observe the chondroprotective mechanism of Tongbi Compound(TC) on adjuvant arthritis(AA) rats.Methods Wistar rats were randomized into the normal group,model group,indomethacin group(2.6 mg?kg-1?d-1),sulfasalazine group(825 mg?kg-1?d-1) and TC group(30 g?kg-1?d-1).Except the normal group,the rats in other groups were given pedal injection of Freund's complete adjuvant(FCA,0.2mg for each rat)to induce AA.After gastric gavage of corresponding medicine for 2 weeks,the pathological changes of the cartilage in the rat ankles and their scores were observed,and the expression of matrix metalloproteinases 3(MMP-3) and tissue inhibitor of metalloproteinase(TIMP-3) in rat cartilage was detected by immunohistochemical method.Results In the model group,pathological score was increased(P0.05),but had less toxic and side effects.Conclusion The therapeutic mechanism of Tongbi Compound for the treatment of ankylosing spondylitis is probably related with the regulation of the balance between MMP-3 and TIMP-3.
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[Objective] To establish the scale of the quality of life for patients with ankylosing spondylitis. [Methods] Initial scale including 58 items was proposed by the research staff. After the preliminary test was performed, 37 items were selected and a confirmed scale of the quality of life for ankylosing spondylitis ( SQOL-AS) was obtained. The feasibility, reliability, validity and sensitivity of SQOL-AS were analyzed. [Results] Statistics showed that SQOL-AS had a good feasibility, reliability, validity and sensitivity. [Conclusion] SQOL-AS can be used to evaluate the quality of life for patients with ankylosing spondylitis.
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[Objective] To establish a method for the determination of brucine and strychnine contents in Tongbiling Tablet. [Methods] Reversed-phase HPLC conditions were: Nucleodur C18 Gravity column (125 mm ? 4.6 mm, 5?m), mobile phrase being methanol-water (60:40) with 0.5 g potassium dihydrogen phosphate and 1.0 g dodecyl sodium sulfouate and 0.5 mL triethylamine in 1 000 mL mixture (adjusting pH to 4.5 with acetic acid) , detecting wavelength at 254 run and a flow rate of 1.0 mL/min at room temperature. [ Results ] The average recovery of brucine was 97.69 % ( RSD = 2.03%) and that of strychnine 97.93% ( RSD = 2.61%). [Conclusion] This method is efficient and can be used for the quality control of Tongbiling Tablet.
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Objective To observe the clinical effect of Erteng Tongbi Mixture(ETM) on rheumatoid arthritis(RA). Methods 102 cases of RA were treated with ETM for 3 months. Time of morning stiffness, rest pain, 20m- walking time, hand grip, the number of joint with swelling and tenderness, erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), C- reactive protein (CRP) were compared before and after the treatment. Results The total effective rate of ETM was 91.4 % and the ineffective rate 8.6 % . The morning stiffness, rest pain, hand grip and joint tenderness were obviously relieved, while 20m- walking time and the number of joint with swelling remained unchanged. ESR, RF and CRP were decreased to different extent, in particular RF(P