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[ ABSTRACT ] AIM: To observe the effect of rapamycin on the apoptosis of mouse astrocytes in vitro.ME-THODS:The astrocytes from C57BL/6J newborn mouse pups were isolated and primarily cultured.The effect of rapamycin on the viability of astrocytes was assessed by MTT assay.The mean fluorescence intensity of SYTOX?Green stain in the astrocytes was detected by fluorescence microplate reader in order to analyze the effects of rapamycin on the cell death in-duced by H2 O2 , ionomycin and/or deferorxamin.DiOC6 (3) staining was used to analyze the mitochondrial membrane po-tential of the astrocytes induced by H2 O2 .Flow cytometry analysis was used to determine the production of ROS in the as-trocytes and mitochondria by staining with H2 DCFDA and MitoSOXTM Red reagent, respectively.RESULTS: Rapamycin at concentration of 0.5 μmol/L protected the astrocytes against cell death induced by H2 O2 or deferoxamine plus ionomy-cin.Rapamycin protected the mitochondrial membrane potential of astrocytes from the injury of H2 O2 .It also reduced the production of ROS in the astrocytes and decreased the level of ROS in the mitochondria.CONCLUSION:Rapamycin re-duces the ROS overload in the mitochondria, keeps mitochondrial membrane potential safety and protects the astrocytes a-gainst apoptosis in vitro.
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BACKGROUND:Because macrophages play an important role in the body’s inflammatory response, the detection of the impact of biological materials on the behavior of macrophages can assess the immunogenicity of materials. OBJECTIVE: To analyze the activation effect of decelularized extracelular matrix materials on macrophages. METHODS: The peritoneal macrophages of BALB/c mouse were obtained and cultured by dividing into five groups. Control group was simple cel culture group, experimental group 1 was acelular matrix membrane material directly contacting with macrophage for culture, experimental group 2 was fresh pericardial material directly contacting with the macrophage for culture, experimental group 3 was acelular matrix membrane material indirectly contacting with macrophages for culture, experimental group 4 was fresh pericardium material indirectly contacting with macrophages for culture. After 24 hours of culture, the secretion of nitric oxide and cytokines in cel culture supernatant was determined. After 48 hours of culture, the absorbance value was determined by MTT method and the toxicity grading was determined. RESULTS AND CONCLUSION: The toxicity grading in experimental groups 1-4 was respectively grades 2, 4, 0, 2. The nitric oxide level in experimental groups 1 and 2 was higher than that in the control group (P < 0.05), and the nitric oxide level in the experimental group 2 was higher than that in the experimental group 1 (P < 0.05). There were no significant differences in interleukin-2, interleukin-4,interferon γ, interleukin-17A and interleukin-10 levels between these five groups. The interleukin-6 level in the experimental group 2 was higher than that in the control group (P < 0.05); The expression levels of tumor necrosis factors in experimental group 1, 2 and 4 were higher than those in the control group (P < 0.05), and experimental group 2 higher than the experimental group 1 (P < 0.05), experimental group 1 higher than the experimental group 4 (P < 0.05). These results show that acelular matrix material can activate macrophages in direct contact.
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Before the technique of advanced high-throughput sequencing comes up , less is known about the human gut microbiota .It has been understood that trillions of microbes , in which 99% are bacteria , inhabit the human gut, forming a complicated ecological community .The gut microbiota has a great impact on human physiology and suscepti -bility to disease through its integrative metabolic activities and interactions with the host .In physiology , gut microbiota con-tributes to the host acquisition of nutrition and energy from diets , promoting development and maturation of gastrointestinal tract and immune system , and protecting host from invasion of enteropathogens .In pathology , dysbiosis underlying altered gut microbiota is associated with the susceptibilities to various diseases , including inflammatory bowel disease , type 1 dia-betes, asthma, obesity, metabolic syndrome , autism and cancer .Understanding of the factors that underlie alterations in the composition and function of gut microbiota will be helpful in the development of drugs and the design of therapies that target it.This goal is formidable .It is because that the compositions of gut microbiota are immensely diverse , varying be-tween individuals in a population and fluctuating over time in an individual , especially during early development and disea-ses.Viewing the gut microbiota with an ecological perspective will provide new insights into how to improve our health by targeting this microbial community in clinical treatments .
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AIM: To investigate the effect of dihydroartemisinin (DHA) on the proliferation of murine T lymphocytes stimulated by Con A in vitro and its related immunosuppressive mechanism. METHODS: Murine T lymphocytes were stimulated by Con A and treated with different concentrations of DHA. Cell proliferation was measured by carboxyl fluoresce in diacetate succinmidyl ester (CFDA-SE) staining. The expression of CD69, CD25 and CD71,which was the marker of early, middle, later activation of CD3~+ T lymphocytes, was measured by flow cytometry (FCM) combined with two-color immunofluorescent staining of cell surface antigen. Fluorescence calcium indicator fluo-4/AM was used to measure the change of the intracellular calcium concentration ([Ca~(2+)]_i) of murine T lymphocytes. The distribution of the cell cycle was analyzed by PI staining. The expression of CD69, the early activation antigen on CD4~+CD25~(high) Treg was also measured by FCM combined with three-color immunofluorescent staining. RESULTS: The result of CFDA-SE staining showed that DHA efficiently inhibited the Con A-induced proliferation of T-lymphocytes in a time-and dose-dependent manners. DHA showed modestly increased proportions of CD69 and CD25 on Con A-stimulated CD3~+T cells, but inhibited the expression of CD25 in a dose dependent manner. DHA with Con A, but not DHA alone, caused an increase in intracellular calcium concentration of T cells. The results of FCM analysis with PI staining showed that DHA imposed a total cell cycle arrest in G_0/G_1 and prevented cells entering S phase and G_2/M phase. Furthermore, DHA reduced the expression of CD69 on CD4~+CD25~(high) Treg. CONCLUSION: DHA, which exhibits immunosuppressive effect on the proliferation of murine T-lymphocytes, is promising to be developed as an immunosuppressive reagent.
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AIM: To investigate the anti-HIV-1 activity of five anthraquinone derivatives (emodin,rhein,chrysophanol,physcion and aloe-emodin) in vitro.METHODS: Viral replication was estimated by observation of cytopathogenesis and measurement of HIV-1 p24 antigen production in HIV-1ⅢB acutely infected C8166 cells. The anti-HIV-1 activity was evaluated by the 50% effective concentrations (EC50) and selective indexes (SI) of these derivatives.RESULTS: These anthraquinone derivatives inhibited HIV-1ⅢB replication on syncytia formation induced by HIV-1ⅢB infection with EC50 mean values of (11.44±0.93)μmol/L (emodin),(51.28±2.86)μmol/L (rhein),(90.58±2.30)μmol/L (chrysophanol),(8.59±0.38)μmol/L (physcion) and (0.89±0.08)μmol/L (aloe-emodin),respectively. The p24 antigen production with EC50 mean values were (11.61±0.56)μmol/L (emodin),(12.35±4.73)μmol/L (rhein),(39.63±2.87)μmol/L (chrysophanol),>250 μmol/L (physcion) and (2.75±0.20)μmol/L (aloe-emodin) respectively. CONCLUSION: These structurally-related chemicals show different anti-HIV-1 activity in vitro. Among them,aloe-emodin is the most potent inhibitor to HIV-1 replication.
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Objective To detect systematic oxidative stress in preeclampsia.Methods (1)Morphological features of placenta hypoxia were observed by histological method ; (2) Level of granulocyte intracellular reactive oxygen species was monitored by dyeing full blood with 2' ,7'-dichlorodihydrofluorescein diacetate (H2DCFDA) ; (3) Level of H_2O_2 in sera was detected by special kits.Results Compared to normal pregnancy,placentas from preeclampsia showed distinct features of hypoxic stress injury,such as more syncytial knots formation,fibrosis emerged,vein in-jury and loss its normal configuration; Fluorescence values of ROS probe in neutrophils from different women were 45.61±12.20(n =49),51.02 ± 13.60(n =56,P <0.01)and 85.10 ± 16.30(n =47,P <0.01); Concentra-tions of H_2O_2were (24.57±5.17)μmol/L(n =49),(26.61±3.25)μmol/L(n =56,P 0.01) and (39.84±9.67)μmol/L(n=47,P<0.01) respectively.Conclusion With the help of histological method,flow cytometry and special kits,systematic oxidative stress can be detected through checking placentic tissues,netrophils and sera of preeclampsia.
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This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.
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BACKGROUND: The connection between Natural killer(NK)-cells and allogeneic bone marrow transplantation(allo-BMT)has aroused increasing attention.OBJECTIVE: To explore the effect of NK cells on graft rejection,hematopoietic and immune reconstitution in mouse undergoing allo-BMT.METHODS: Lethally and nonlethally irradiated BALB/c(H-2d)mice were transplanted with C57BL/6(H-2b)bone marrow plus donor peripheral T cells and/or NK cells.RESULTS AND CONCLUSION: Compared with lethally irradiated and allo-BMT group without infusion of NK cells,the survival rate in lethally irradiated and allo-BMT group with infusion of NK cells significantly enhanced; leukocytes count,expression level of CD19+and CD34+cell count recovered rapidly; expression level of H-2b*cell obviously increased.Expression level of CD34"cell in the group with infusion of NK cells was obviously lower than that of the group without infusion of NK cells at 28 days after transplantation,but there was no significant difference between the 2 groups at 60 days(P > 0.05).In nonlethally irradiated and allo-BMT group without NK cell infusion,expression level of H-2b*cell significantly decreased at 30 days after transplantation,and reduced to before transplantation level at 60 days; while expression of H-2b+cell yet could be detected with more than 80% at 60 days after transplantation in group infused with high and low concentration of NK cells.In alIo-BMT mice,alloreactive NK cell inhibits graft rejection,enhances engraftment,promotes the reconstitution of hematopoiesis and immunity,and increases survival rates.
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This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.
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BACKGROUND: CpG oligodeoxynucleotide (ODN) is a type of highly effective immune adjuvant with low toxicity, which has an extensive application in gene therapy for many diseases. However, the specificity for species and cells leading to low uptake by cells and degradation by nuclease blocks its clinical application. OBJECTIVE: To explore the specific delivery and its immunologic efficacy of CpG ODN targeting B lymphocytes of umbilical cord blood by CD40 ligand-receptor-mediated carrier system. DESIGN, TIME AND SETTING: An observation and control experiment was performed at the Department of Hematology, and Department of Pediatric, the First Affiliated Hospital of Jinan University from April 2004 to October 2007. MATERIALS: Fresh umbilical cord blood with heparin was obtained from healthy, natal infant. Informed consent was obtained from his parents, and the experiment was approved by the hospital Ethics Committee. METHODS: CD40 ligand (CD40L)-EDC-PLL-CpG ODN conjugated complex was prepared. Mononuclear cells (MNCs) from umbilical cord blood were co-cultured with conjugated complexes. Uptake rate, mean fluorescence intensity of FAM marked CpG ODN, expressions of MNCs, proliferations of lymphocytes and the IgG levels of culture supematants were detected by flow cytometry, fluorescence techniques, MTT assay and ELISA, respectively. MAIN OUTCOME MEASURES: The uptake rate, the mean fluorescence intensity of CpG ODN by MNCs, subgroups and proliferations of lymphocytes, and IgG levels of culture supematants. RESULTS: Compared to the pure CpG ODN group, the uptake rate of the conjugated complexes group was higher (98%), the peak level of up-taking occurred earlier, and intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, and CD20+ was increased, A value and IgG levels in supematants were all higher than that of the control group. CONCLUSION: CD40 tigand-receptor-mediated carrier system is helpful for CpG ODN delivery targeting to B lymphocyte, enhancing its immunological efficiency.
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Objective To investigate effect of first, second, and third trimester placental factors (PF) on CD4, CCR5, and CXCR4 expression in human peripheral blood lymphocytes (PBLs), and to explore their influence on human immunodeficiency virus (HIV) vertical transmission.Methods Human peripheral blood mononuclear cells (PBMCs) were treated with first, second,and third trimester PF (concentration 25%) respectively for 24 hours. The expression of CD4, CCR5,and CXCR4 in PBLs, and the percentages of CCR5+, CXCR4+,and CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes were determined with flow cytometry.Results All trimester PFs reduced CCR5 expression in PBLs. The efficiency of the first trimester PF was higher than that of the second and third trimester PF. The percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of PF groups was significantly lower than that of the control group, and the percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the second and third trimester group. The percentages of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of PF groups were significantly decreased as compared with the control group, and the percentage of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the third trimester PF group.Conclusion PF can reduce the expression of CCR5 in human PBLs and peripheral blood CD4+ lymphocytes, indicating that PF might reduce R5 virus infection via preventing HIV entry, and might play an important role in reducing R5 virus intrauterine infection.
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The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide(NO) secretion of mouse macrophages stimulated by lipopelysaccharide (LPS) for 24 h was assayed by Griess reagent system. We found that RCE had potent anti-inflammatory effects on mice. RCE had littl ecell toxic effect on mouse lymphocytes and macrophages. RCE strongly inhibited the excessive production of inflammatory mediators ( NO, CD69, CD25, CD71 ), in a dose-dependent manner, like cyclosporine A injection. RCE could inhibit proliferation of CD3<'+> T lymphocytes. These data suggested that RCE migh texhibit anti-inflammatory effect by inhibiting the activation and proliferation of mouse lymphocytes and the NO secretion of mouse macrophages.
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Objective To investigate the T cell cytotoxicity induced by recombinant adenovirus carrying HIV-1 vpr gene.Methods C8166 cells infected with rAd-vpr or negative control rAd-vector,were analyzed for cell cycle distribution and cell death by flow cytometry.The discrimination of living cells,apoptotic and necrotic cells were differentiated with Hoechst-PI double staining under the confocal microscopy.Changes of mitochondrial membrane potential(△ψm)were monitored by JC-1 staining method.Results Annexin V-PI and Hoechst-PI staining indicated the death effects of HIV-1 Vpr on C8166 cells.PI flow cytometric analysis showed that cell cycle arrested in G2 phase.C8166 cell△ψm collapse mediated by Vpr was detected by JC-1 fluorescent staining.Conclusion The ability of recombinant adenovirus carrying HIV-1 vpr gene to induce mitochondria dysfunction,cell cycle G2 phase arrest and cell death was confirmed in C8166 cells.
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Objective To investigate the effects of protein kinase C (PKC) on proliferation, extracelluar matrix synthesis and transcriptional factor SOX9 (SRY-related high mobility group-box gene 9) expression of rat growth plate chondrocytes in vitro. Methods Rat costochondral growth plate chondrocytes (RGC) were isolated and cultured. The 1st serum free cultured passage RGCs were treated with 1, 10 and 100 nmol/L phorbol 12,13-dibutyrate (PDBu), PKC agonist, cell morphology were observed with inverted microscopy, cell proliferation, COLLAGEN and GAG synthesis were detected by isotope incorporation COLLAGEN TYPEⅡ and AGGRECAN mRNA transcription and SOX9 expression were revealed by RT-PCR and Western blot.Results 100 nmol/L PDBu treatment made the cell morphology of serum free cultured RGC closed to serum group and inhibited proliferation but promoted COLLAGEN and AGGRECAN synthesis, 3H-TdR、3H-proline and 35S-sulfate incorporation of 100nmol/L PDBu group were as 83%, 52.6% and 146.5% as those of serum free control(P
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AIM: To investigate the potential of murine epidermal stem cell (ESC) differentiation after seeded in a biodegradable carrier and implanted subcutaneously into syngeneic recipient mice. METHODS: ES cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a polyglycolic acid (PGA) net containing collagen gel, functioning as a cell carrier, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. RESULTS: The ESCs kept alive in the implant when observed under a fluorescent microscopy 3 weeks or longer after implantation, and could differentiate into hair follicle - like structure,glandular structure, and gave rise to additional structures displaying features resembling native dermis. No apparent rejection or severe side effects were observed at least 10 weeks post- implantation. CONCLUSION: It is feasible to use these ESCs as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.
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Objective: To observe the effect of Trifoliumpratense Leguminosae extract (TLE) on mouse allogenetic skin transplantation. Methods: Recipient BALB/c was divided into physiologic saline (PS) group and TLE group, full-thickness skins were transplanted through back to back method from donor C57BL/6. The allogenetic transplanted skin growth condition was observed. The proliferation of lymphocytes of recipient mice in vitro were detected by CFDA-SE stain and mixed lymphocyte reaction respectively. Results: The allogenetic transplanted skin injected with TLE 25g/kg per day by vena caudalis growed better than that in PS group. The proliferation of lymphocyte in TLE group was smaller than that in PS group. Conclusion: TLE maybe participate in the regulation of mouse immune system and induce its tolerance to the allogenetic transplanted skin.
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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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AIM To investigate the effect of recipient kidney function by CsA coadministration Ber used to induce immune tolerance in rats of allogenic cardiac transplantation. METHODS The authors established the SD to Wistar rats heterotopic cardiac transplantation model by Onos methods.Observe the cardiac allograft survival and levels of BUN and Cr in the recipients plasma. The recipients were classified into 5 groups randomly after heterotopic cardiac transplantation were performed. Group A (Wistar to Wistar)): Received placebo intraperitoneal injected for 21 days; Group B (SD to Wistar): Saline intraperitoneal injected for 21 days; Group C (SD to Wistar):CsA 2 mg?kg -1 ?d -1 intraperitoneal injected for 21 days; Group D(SD to Wistar):Ber 16 mg?kg -1 ?d -1 gastrointubation for 21 days; Group E(SD to Wistar): Ber 16 mg?kg -1 ?d -1 gastrointubation coadministration CsA 2 mg?kg -1 ?d -1 ip for 21 days. RESULTS The levels of BUN and Cr in recipint plasma is lower evidently compare with the group with CsA ip simply. CONCLUSION Ber can reduce the renal toxicity in recipients by CsA which was intraperitoneal injected (ip) over a long period time.
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Aim To investigate effects of genistein (5, 7, 4′-trihydroxyisoflavone) on rat costochondral growth plate chondrocyte (RGC) collagen and Sox9 expression. Methods Primary cultured RGC, effects of genistein on collagen synthesis, col2a1 mRNA transcription and Sox9 protein expression of 1st passage RGC were detected by isotope incorporation, RT-PCR and western blotting. Results Genistein inhibited collagen synthesis of 1st passage RGC (P