BACKGROUND: The main role that the osteoprotegedn (OPG) plays in bone tissues is to inhibilate the formation and the activity of osteoclast, while as for the receptor activator of nuclear factor-κB ligand (RANKL), it is to stimulate the differentiation and the activity of osteoclast. Both OPG and RANKL are important for the regulation of osteoclastic function. Recent studies have found the stimulative effects of adiponectin on the proliferation and the differentiation of osteoblast, as well as the coupling between adiponectin and bone metabolism. But effects of adiponectin on osteoclast remain unclear. OBJECTIVE: To investigate the effect of adiponectin on the expression of OPG and RANKL in osteoblast, and to further analyze its effects on the formation of osteoclast. DESIGN, TIME AND SETTING: A contrast observational experiment was conducted in the laboratory of Institute of 2008.MATERIALS: Cancellous bone in anterior superior lilac spine was obtained from adult normal by surgery for cell incubation. Clinical samples were provided by the Xiangya Hospital. METHODS: Human osteoblast was incubated with different doses of adiponcetin (0, 3, 10 and 30 mg/L) for 48 hours, after which OPG and RANKL mRNA and protein expression level was determined. In addition, adiponcetin was added into the co-culture system of osteoblast and peripheral blood monouclear cells for examing its effects on osteoclast formation. MAIN OUTCOME MEASURES: OPG and RANKL mRNA and protein expression in human osteoblast was determined using fluorescent quantitative polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA). Osteoclast was detected by antitartaric acid acid phosphatase staining. RESULTS: Adiponcetin inhibited osteoblast OPG mRNA and protein expression in a dose-dependent way (P < 0.05). Adiponcetin promoted osteoblast RANKL mRNA and protein expression in a dose-dependent way (P < 0.05). Adiponectin induced the osteoclasts formation in the co-culture system of osteoblast and peripheral blood monouclear cells. CONCLUSION: Adiponectin has the effect of inducing osteoclast formation via stimulating RANKL expression and inhibiting OPG

Objective To investigate the influence of indomethacin to the unite treatment effect on chronic phase myeloid leukemia (CML-CP) with interferon alpha-2b (IFNα-2b) and low dose cytarabine (LD-Ara-C).Methods 22 CML-CP patients were randomly divided into two groups.Control groups (10 cases) injected with IFNα-2b (3 million units),injection frequency was q.o.d,the duration of treatment was about 3-18 months,cytarabine (Ara-C) by slowly intravenous driped (30 mg/d).In this treatment schedule,every course of treatment sustained 10 days,and with a 2 weeks interval.During this process,patients in treatment group were treated with hydroxyl urea only when their WBC in peripheral blood exceed 20×109/L,otherwise,discontinue it.Treatment group (12 cases),on the first day of treatment with IFNα-2b and Ara-C,added indomethacin (25 mg) through oral administration,the frequency was t.i.d.During treatment in the two group,the end point of observation was completely hematology ease,at the same time,these indicators in the two groups needed to be compared,the time when WBC begin to fall,the time when WBC fall to normal range,the time when immature cells returned to normal,the time which complete hematological remission and the highest temperature of patients after IFNα-2b was subcutaneous injected.Results The time when WBC begin to fall in treatment group was (4.2±2.7) d,and the time was (5.0±2.5) d in control group (t =0.714,P ＞ 0.05).In treatment group,the time when WBC fall to normal range was(10.0±4.5) d,and the time was (12.0±4.5) d in control group (t =1.036,P ＞ 0.05).The time when immature cells returned to normal in treatment group was (14.2±4.8) d,and the time was (19.0±3.6) d in control group (t =2.609,P ＜ 0.02).The time which complete hematological remission was achieved in control group was (45.8±5.6) d,but it was (53.9±10.5) d in control group (t =2.314,· P ＜ 0.05).Meanwhile,the fever degree after IFNα-2b was subcutaneous injected obviously achieved improvement in treatment group (x2 =12.041,P ＜ 0.005).Conclusion The advantage of indomethacin to the unite treatment with IFNα-2b and LD-Ara-C on CML mainly lays in which cound alleviated the adverse reaction such as flu-like of IFN,and more,there are synergy effect in antagonist CML.

Objective To evaluate the effect of curcumin on the proliferation of and apoptosis in HaCaT cells induced by tumor necrosis factor α (TNF-α).Methods HaCaT cells were cultured with the presence of different concentrations (0,1,5,10,25,50,100 ng/ml) of recombinant TNF-α,curcumin of 20 μmol/L,or the combination of recombinant TNF-α (25 ng/ml) and curcumin (20 μmol/L),for 24 hours followed by the determination of cell proliferation with methyl thiazolyl tetrazolium (MTT) assay.Western blot was conducted to measure the protein expression of proliferating cell nuclear antigen (PCNA) and Notch-1 in HaCaT cells treated with recombinant TNF-α (25 ng/ml) and curcumin (20 μ mol/L) alone or in combination for 24 hours.Flow cytometry using annexin-V/propidium iodine (PI) was performed to assess the early apoptosis in HaCaT cells incubated with recombinant TNF-α of 25 ng/ml and curcumin of 20 μmol/L alone or in combination for 12 hours.Statistical analysis was carried out with one-way analysis of variance.Results Recombinant TNF-α promoted the proliferation of HaCaT cells in a dose-dependent manner,with the maximum proliferation activity observed in HaCaT cells treated with TNF-α of 25 ng/ml,while curcumin of 20 μmol/L effectively inhibited the proliferation of HaCaT cells induced by TNF-α of 25 ng/ml (P ＜ 0.01).TNF-α of 25 ng/ml had no obvious effect on cell apoptosis,while curcumin of 20 μ mol/L markedly induced the apoptosis in HaCaT cells,and there was a synergy between TNF-α of 25 ng/ml and curcumin of 20 μmol/L in the induction of apoptosis in HaCaT cells,with the apoptosis rate being 2.3％,3.4％,11.6％ and 16.8％ respectively in untreated cells,cells treated with TNF-α,curcumin,and the combination of TNF-α and curcumin,respectively.Conclusions Curcumin could enhance the inductive effect of TNF-α on the apoptosis in,but suppress the promotive effect of TNF-α on the proliferation of,HaCaT cells.

Purpose　To study the main points of the diagnosis and differential diagnosis of the lymphadenitis in CSD. Methods　Expression of CD45, CD3 and CD68 were evaluated immunohistochemically (S-P method). Results　Five cases of microabscess in early stage and 21 cases of microabscess-granuloma were found. The histologic features were the formation of microabscess and granuloma. The early microabscess in the lymphnodes were surounded by B lymphocytes (CD20+) and macrophages (CD68+). The typial microabscess granuloma were surrounded by epitheloid cells (CD68+) and CD3 positive T lymphocytes. Conclusion　The main characteristic of the lymphadenitis of cat scratch disease is the formation of granuloma with microabscess. The immumohistochemical markers are useful to distinguish the proliferative cell types. This lesion may result from bacterial infection which induces the cell immune reaction.

Models of throat excess-heat mice were established and different dosages of water extract of STRR were ig given to mice to observe anti-inflammatory effect and its mechanism. The activities of ALT, AST and the contents of TNF-α, T3, rT3, T4, SOD, MDA, PEG2, NO, NOS, Cr, BUN, GSH and GSH-Px in serum were tested while liver index, kidney index, spleen index and thymus index were measured. The anti-inflammatory efficacy accompanied by side effects and mechanisms of water extract of Sophorae Tonkinensis Radix et Rhizoma (STRR) in excess-heat mice were investigated to clear safety dose-dependence range and the relationship of efficacy, toxicity and syndrome. In the experiment, water extract of STRR showed a strong inhibitory effect on ear edema by croton oil in throat excess-heat mice. The activities of ALT, AST in serum and liver index were all higher than that of normal group after multiple administration. PEG2, SOD, MDA, NO, NOS, GSH and GSH-Px had obvious changes. According to the results, water extract of STRR has an anti-inflammatory effect on acute inflammation in throat excess-heat mice and it is stronger than that in normal mice. The anti-inflammatory effect of STRR is related to the reduction of inflammatory mediators release. Side effects and hepatotoxicity will be produced on clinical efficacy dosage. The mechanisms of anti-inflammation and hepatotoxicity are all in connection with oxidative damnification.
Animals ; Anti-Inflammatory Agents ; therapeutic use ; Chemical and Drug Induced Liver Injury ; etiology ; Disease Models, Animal ; Female ; Male ; Medicine, Chinese Traditional ; Mice ; Phytotherapy ; Plant Extracts ; therapeutic use ; toxicity ; Rhizome ; Sophora

BACKGROUND: The correlation of gycosylphosphatidilinoditol-specific phospholipase D (GPI-PLD) activity, mRNA expression to leukemia type, hepatosplenomegaly and/or lymphadenopathy has been rarely reported. OBJECTIVE: To explore the correlation of GPI-PLD expression to leukemia type and hepatosplenomegaly and/or lymphadenopathy of acute myeloid leukemia (AML) patients. METHODS: Fresh bone marrow specimens were obtained from 43 newly diagnosed AML patients, 28 acute lymphocytic leukemia (ALL) patients, and 21 normal persons. Bone marrow mononuclear cells were harvested by density gradient centrifugation. GPI-anchored human placent alkaline phosphatase was used as substrate. GPI-PLD activity was determined bytriton-X114 phase partitioning procedure. GPI-PLD mRNA expression was detected by semi-quantitative RT-PCR. The relationship of GPI-PLD activity, mRNA expression and leukemia type, hepatosplenomegaly and/or lymphadenopathy was analyzed. RESULTS AND CONCLUSION: Compared with control group, GPI-PLD activity and mRNA expression in bone marrow mononuclear cells were significantly higher in AML group (P < 0.01), while they were significantly lower in the ALL group (P < 0.01). Of 43 patients with AML patients, 13 patients had hepatosplenomegaly and/or lymphadenopathy. The GPI-PLD activity (%) and mRNA expression were significantly higher in AML patients without hepatosplenomegaly and lymphadenopathy than those patients with hepatosplenomegaly and/or lymphadenopathy (P < 0.05). These results demonstrated that GPI-PLD activity alteration is consistent with GPI-PLD mRNA expression in AML patients, and the expression levels correlate to leukemia type and hepatosplenomegaly and/or lymphadenopathy of AML patients.

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

OBJECTIVETo explore efficacy enhancing and detoxification roles of Jiedu Quyu Zishen Recipe (JQZR) in treating systemic lupus erythematosus (SLE) by studying its effect on Toll like receptor 9 (TLR9) signal pathway of murine macrophage cells after JQZR stimulated CpG oligodeoxynucletide (CpG ODN).

METHODSMurine macrophage cells in vitro cultured were randomly divided into 4 groups, i.e., the blank serum group, the CpG ODN stimulus group, the CpG ODN + dexamethasone group, the CpG ODN + medicated serum group. Murine macrophage cells were collected after 24-h intervention. The expression of TLR9, myeloid differentiation factor 88 (MyD88), NF-KB, IFN-α mRNA were analyzed by RT-PCR. The expression of TLR9 and NF-κB protein were analyzed by Western blot. Changes of the NF-KB transcriptional activity were assayed by Dual-Luciferase reporter assay system.

RESULTSmRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were enhanced, showing statistical difference when compared with those of the blank serum group (P <0. 05, P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of MyD88, NF-κB, and IFN-α, the protein expression of NF-κB and the NF-κB transcriptional activities decreased in the CpG ODN + dexamethasone group with statistical difference (P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were decreased in CpG ODN+ medicated serum group with statistical difference (P <0. 01).

CONCLUSIONEfficacy enhancing and detoxification roles of JQZR in treatment of SLE might be realized through regulating TLR9 signal pathways.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Macrophages ; metabolism ; Mice ; Myeloid Differentiation Factor 88 ; NF-kappa B ; RNA, Messenger ; Signal Transduction ; Toll-Like Receptor 9 ; metabolism

Objective To explore the interaction between serum melatonin and the status of disease and probe the effect factor of serum melatonin change in asthmatic children.Methods Serum melatonin was measured in asthmatic children with 15 cases of mild persistent asthma,15 cases of moderate persistent asthma,15 cases of severe persistent asthma,15 cases of stable asthma and 15 cases of normal subjects by enzyme-linked immunosorbent assay(ELISA).Results The levels of serum melatonin in the 5 groups of mild persistent asthma,moderate presistent asthma,Severe Persistent asthma,Stable asthma,control subject were(22.76?5.16)ng/L,(16.79?3.35)ng/L,(11.54?1.45)ng/L,(22.06?3.36)ng/L,(28.72?4.32)ng/L,respectively.There were significant differences between any of them(Pa

Objective To report a case of the implantation of thyroid hyperplastic or neoplastic tissue after endoscopic thyroidectomy and discuss this complication in aspects of prevalence, pathogenesis, protection, and therapies.
Methods A systematic search of literature from the PubMed database was conducted for identifying eligible studies on implantation of thyroid hyperplastic or neoplastic cells after endoscopic thyroid surgery.
Results Overall, 5 reported cases on patients suffering from endoscopic thyroid surgery with implantation of thyroid hyperplastic or neoplastic cells were included in the systematic review.
Conclusions Unskilled surgeons, rough intraoperative surgical treatment, scarification or rupture of tumor, contamination of instruments, chimney effect, aerosolization of tumor cells may be associated with the implantation after endoscopic thyroidectomy. To minimize the risk of such complication, we should be more meticulous and strict the endoscopic surgery indications.