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Objective To explore the value of MR T2 mapping imaging in evaluation of the effection of Zhitong-Jiangu recipes in treatment of cartilage injury in osteoarthritis.Methods The models of osteoarthritis in the knee of New Zealand white rabbits were made with intra-articular injection of papain.Totally 48 New Zealand white rabbits were randomly divided into normal control group,model control group,glucosamine hydrochloride treatment group and Zhitong-Jiangu recipes treatment group.MR T2 mapping was performed on the knee and the content of MMP-1 in articular cartilage was analyzed with immunohistochemistry 4 weeks after treatment.The differences of T2 value and MMP-1 content of articular cartilage among the four groups were compared.The correlation among the different intervention methods,MMP-1 content of articular cartilage and the T2 value of articular cartilage were analyzed.Results T2 value of articular cartilage in the model control group was significantly higher than that in the other three groups (all P＜0.01),and there was no significant difference of T2 value among the other three groups.The MMP-1 content of articular cartilage in glucosamine hydrochloride treatment group and Zhitong-Jiangu recipes treatment group was lower than that in model control group (both P＜0.05),and was higher than that in normal control group (both P＜0.05).There was no significant difference between the two groups.The relationship between MMP-1 content and T2 value of articular cartilage was nonlinear.With the increase of MMP-1 content,T2 value first increased slowly and then increased rapidly (P＜0.05).Conclusion MR T2 mapping imaging is helpful to evaluating the efficacy of Zhitong-Jiangu recipes in treatment of cartilage injury in osteoarthritis in rabbit models.The relationship between MMP-1 content and T2 value of articular cartilage may be nonlinear.
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Objective:To explore the effect of Jinfeng particles on the expression of MMP-1/TIMP-1 in cartilage of gout arthritis rabbit′s articular,to explicit treatment mechanism of gouty arthritis. Methods:Perfusion of ethambutol and adenine,inject MSU into joint cavity of rabbits to copy the model of gouty arthritis. After successful mode,were randomly divided into model group,Jinfeng granules in high,medium and low dose group(19. 6,9. 8,4. 9 g/kg),allopurinol group(0. 028 g/kg),Tongfengding group(0. 448 g/kg) ,another blank group. Administered for 7 days. 2 h after the last administration,were taken and painted articular cartilage tissue,to observe the changes of the pathological morphology;and the use of immune cell detection of the expression of TIMP-1,MMP-1,MMP-1/TIMP-1 ratio calculation. Results: Jinfeng granule could obviously reduce the articular cartilage of rats with inflammatory cells infiltration,hyperemia, swelling and necrosis, could significantly inhibit the expression of MMP-1 ( P< 0. 05 ) , stimulate TIMP-1 activation(P< 0. 01),and significantly reduce the ratio of MMP-1/TIMP-1(P< 0. 05). Conclusion: Jinfeng particles can effectively inhibit the pathological damage of gouty arthritis,upside the expression of TIMP-1 and its mechanism may be related to the inhibition of MMP-1,regulated the balance of MMP-1/TIMP-1,and then exert anti-inflammatory effects.
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Objective To observe the influencing of Yiqi Huayu Jiedu Prescription on the growth of HepG2 nude mice transplantation tumor and the expression of related factors of vascular mimicry. Methods Models of transplanted tumors, which were made by HepG2 cells in nude mice, were randomly divided into 7 groups, Yiqi Huayu Jiedu Prescription group, Astragali Radix group, Curcumae Rhizoma group, Paridis Rhizoma group, Gecko group, cis-platinum group, and model group. Except for the model group, the rest groups were given relevant medicine for intervention. 21 days later. HIF-1α, MMP-2, MMP-9, and E-cad were detected by immunohistochemistry, and Twist1 and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the model group, tumor volume in the rest groups decreased (P<0.05), and the effect in the Yiqi Huayu Jiedu Prescription group was more obvious than the Astragali Radix group, Paridis Rhizoma group and Gecko group (P<0.05);The expression of vasculogenic mimicry structure was rare in each group, and the model group and cis-platinum group were the most obvious;Except for the Astragali Radix group, the expressions of HIF-1α, MMP-2, and MMP-9 showed statistical significance compared with model group (P<0.05);The expression of E-cad in the Yiqi Huayu Jiedu Prescription group and Astragali Radix group showed statistical significance (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group, Paridis Rhizoma group, and Gecko group decreased significantly compared with the model group (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group was much better than the other groups (P<0.05);The expression of Twist1 showed statistical significance in the Yiqi Huayu Jiedu Prescription group, Curcumae Rhizoma group, Paridis Rhizoma group, and cis-platinum group (P<0.05). Conclusion Yiqi Huayu Jiedu Prescription can reduce expression of HIF-1α, Twist1, Bcl-2, MMP-2, and MMP-9, and increase expression of E-cad, thereby inhibiting the formation of vascular mimicry.
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Objective To evaluate the diagnostic value of immunohistochemistry (IHC) for the identification and localization of Treponema pallidum (TP).Methods Rabbit anti-human TP polyclonal antibody labeled IHC was used to detect 20 paraffin-embedded biopsy samples from lesions of 14 patients with syphilis or suspected syphilis in Sir Run Run Shaw Hospital of Zhejiang University from January 2004 to May 2012.Results TP was detected in 80％ of all the 20 samples by IHC assay,including 83.3％ (5/6) in patients with primary syphilis,100.0％ (10/10) in patients with secondary syphilis,and 25.0％ (1/4) in patients with tertiary syphilis,with a positive diagnostic accuracy of 100.0％.TP was mainly present in lower part of epidermis or perivascular,characterized by an endotheliotropic and epitheliotropic patterns or in the tissue of granulomatous inflammation.Besides,the density of TP was associated with types of lesions.There were more TP in the lesions of syphilis chancre,syphilis proctitis and condyloma latum,and fewer TP in the lesions of squamous erythema,greyish-black plaque,ulcer of chest wall from tertiary syphilis,and least in syphilitic lymphadenitis.There were no correlations between the quantity of TP and the rapid plasma regain (RPR) test titer (P＞0.05).Conclusions IHC for TP is of both high sensitivity and specificity for the diagnosis of syphilis,suggesting that TP-IHC is helpful for the diagnosis of syphilis,especially for the diagnosis of early suspected syphilis with negative serological results,systemic damage of syphilis,or syphilis in untypical locations and unusual lesions.It can serve as an alternative method for the diagnosis of syphilis.
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<p><b>OBJECTIVE</b>To investigate the effect of triptolide on the differentiation of human Th17 cells.</p><p><b>METHOD</b>Human peripheral blood mononuclear cells, purified CD4+ T cells and CD4+CD45RA- memory T cells were treated with various concentrations of triptolide in vitro. Cell proliferation was determined by MTT assay. Flow cytometry was used to analyze the intracellular expression of IL-17 and IFN-gamma. Cytokine production of IL-17 and IFN-gamma was measured by ELISA.</p><p><b>RESULT</b>Cell proliferation, intracellular expression of IL-17 and IL-17 secretion were inhibited by triptolide in a dose-dependent manner. IFN-gamma expression and production were also inhibited by triptolide.</p><p><b>CONCLUSION</b>Triptolide inhibits the differentiation of human Th17 cell. The observation may indicate at least one of the mechanisms of the immunosuppressive and anti-inflammatory effects of triptolide.</p>
Subject(s)
Humans , Anti-Inflammatory Agents, Non-Steroidal , CD4-Positive T-Lymphocytes , Metabolism , Cell Differentiation , Cell Proliferation , Cells, Cultured , Diterpenes , Pharmacology , Dose-Response Relationship, Drug , Epoxy Compounds , Pharmacology , Interferon-gamma , Metabolism , Interleukin-17 , Metabolism , Bodily Secretions , Phenanthrenes , Pharmacology , Th17 Cells , Cell Biology , Tripterygium , ChemistryABSTRACT
Objective To investigate the expression of Toll-like receptors(TLRs) in condyloma acuminatum(CA) lesions and their possible roles in the pathogenesis of CA. Methods The expressions of TLR1-10 mRNA level in the lesions of CA and in the cervix scrape cells from the patients with human papillomavirus(HPV) negative chronic cervicitis were detected by real-time quantitative fluorescent PCR. HPV typing was detected by HPV GenoArray test kit. Results Low-risk HPV type 6 and type 11 were the most prevalent types in the forty CA cases with positive rate of 77.5% and 55% respectively. 55% CA patients were found infected with more than two types of HPV. 35% CA patients were concurrently infected with high-risk HPV. The expressions of TLR3, 7, 8 mRNA were higher than other TLRs and the expression of TLR9 mRNA was lower than others in the lesions of CA. No significant differences of the TLR1-10 mRNA levels were found between HPV6 and HPV11 positive CA lesions, so did it between low-risk and high-risk HPV concurrent infected CA lesions. The expressions of TLR1-3, TLR5-8, TLR10 mRNA, especially TLR2, TLR7 and TLR8 in the lesions of CA were significantly higher than that in cervix scrape cells of HPV negative chronic cervicitis. There were no significant differences of TLR4 and TLR9 mRNA levels between the two groups. Conclusion There were higher expressions of some TLRs (3, 7, 8) and lower expression of TLR9 in the lesions of CA. Compared with HPV negative chronic cervicitis, the expressions of TLR1-3, TLR5-8, TLR10 mRNA in the lesions of CA were up-regulated. The expression profile of TLRs in different type of HPV infected CA lesions had no significant differences. Our results suggested that the expression profile of TLRs in CA may be associated with the HPV infection. Whether it was associated with the immune escape mechanism and persistent infection of HPV need further demonstration.
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Objective To observe the levels of Foxp3+ CD+ CD25+ regulatory T cells in the peripheral blood of condyloma acuminatum (CA) patients and investigate their roles in the pathogenesis of CA. Methods The peripheral blood was collected from 30 CA patients (including 15 with relapsing and 15 with first onset) and 20 healthy controls. Peripheral blood mononuclear ceils (PBMC) were isolated and stained with anti-human CD4-PE-Cy5 and anti-human CD25-fluorescein isothiocyanate (FITC) monoclonal antibodies on cell membrane, followed by intraeellular staining with anti-human Foxp3-PE. The percentage of Foxp3+ CD4+-CD25+ regulatory T cells was detected by three-color flow cytometry. Comparison between groups was done by ANOVA test. Results The percentages of Foxp3+ CD4+ CD25+ regulatory T cells among total CD4 + T cells in CA patients and relapsing CA group were (3.4 ± 1.0) % and (4.7 ±+ 1.2) %, respectively, which were both significantly higher than that in healthy control group [(1.2±0. 5)%, P<0.01]. Furthermore, that in first onset CA group was (2. 1 ± 1.0) %, which was higher than that in healthy control group, but without statistical significance; but that in relapsing CA group was significantly higher than that in first onset group (P<0.05). Conclusions The number of Foxp3+ CD4+ CD25+ regulatory T cells increases in the peripheral blood of CA patients. The disorder of cellular immunity may be involved in the immunopathogenesis of CA.
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We put forward that ①Etiological factors,pathogenesis and manifestations of a disease must be carefully studied before a syndrome differentiation standard is set up;②The ratio equilibrium of disease and syndrome should be considered when grouping in the process of a study;③In the process of designing a experiment,cases with similar symptoms or contradictory symptoms to the study cases are chosen to the control group;④Similar drug or two drugs used in combination should be adopted for the control drug in case of no same kind drug avalable.
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Objective To screen and identify the predicted epitopes of synthesized HLA-A*0201restricted CTL derived from HPVll E7 antigen.Methods Five HPVll E7 CTL epitope peptides and terramers consisting of HLA-A*0201 were selected by way of computer and synthesized by Sanquin company,including HPVllE7 7-15(TLKDIVLDL),15-23(LQPPDPVGL),47-55(PLTQHYQIL),81-89(DLLLGTLNI)and 82-90(LLLGTLNIV).These peptides binding to human peripheral blood-derived DCs were tested for their ability to activate T cells isolated from peripheral blood lymphocytes of HLA-A*0201 healthy individuals.the number of specific tetramer+CD8+T cells by flow cytometry,the level of the section of IFN-γ by ELISA,and the ability of the CTL to kill the target cells were observed.Results The immature DCs could be fully activated by all the five HPV11 E7 peptides.Peptide-loaded mature DCs were able to stimulate the epitope-specific T cells responses in vitro.An increased frequency(P<0.05)of T ceils specific for the E7 7-15 epitope compared to other epitopes of HPV11E7.The epitope-specific CTL of E7 7-15 induced by the activated DCs specifically killed HPV11E7 expressing 293 cell line,and in a ratio of 50:1,the specific cytolytic activity was the strongest than the others(P<0.05).Conclusion DCs loaded with HPV11 E7 7-15(TLKDIVLDL)peptide can induce highly effective and specific ectogenic processed epitopespecific CTL responses in vitro.This peptide may be the candidate for development of CTL based vaccine in the treatment of HPV infeetions.
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Objective To approach the main symptoms and pulses of excessive rising of live yang in hypertensive disease. Method The symptoms and pulses of excessive rising of liver yang in hypertensive disease were collected through clinical epidemiology. The frequency of single symptom and pulse or their combinating group was statistical analyzed by multiple factor cluster analysis and principal component analysis. Result Such symptoms and pulses as lightheadedness,headache,distention of head,tantrum,insomnia,dizzy,bitter taste of mouth,red tongue,yellow tongue fur and string pulse not only can reflect the main pathogenesis,but also are the most frequently occurring reflection. Conclusion Symptoms and pulses above are the differentiation of symptoms and signs standard for excessive rising of liver yang in hypertensive disease.
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Objective To elucidate the therapeutic mechanism of 1,25-dihydroxyvitamin D3 and its analogues in psoriasis. Methods The modulating role of calcitriol, calcipotriol and tacalcitol on the expression and secretion of IL-10 in monocyte-derived dendritic cells (MDDCs) was investigated. Human monocytes were differentiated into MDDCs in vitro in the presence of GM-CSF and IL-4. Semi-quantitative RT-PCR and ELISA were used to analyze the mRNA expression and protein production of IL-10 in MDDCs. In each experi-ment, 24,25-dihydroxyvitamin D3 (a vitamin D3 metabolite with a low affinity to the vitamin D receptor) was used as a control. Results The mRNA expression and protein secretion of IL-10 in MDDCs was up-regulated by calcitriol, calcipotriol or tacalcitol with a similar potency, but not by 24,25-dihydroxyvitamin D3. The up-regulatory role of calcitriol on IL-10 protein secretion was dose dependent. Conclusion The therapeutic effect of calcitriol and its analogues in the treatment of psoriasis may be related to the induction of IL-10 production.
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Objective To explore the methods of large sample research for traditional Chinese medicine(TCM) symptoms of exercise-induced diseases and to define the main symptoms and the main symptom compositon of exercise-induced diseases.Methods The methods of TCM theory of differentiation of syndrome, epidemiology study and multivariable statistical analysis were properly combined. We investigated 518 cases of exercise-induced insomnia(EII) by using the clinical epidemiology program with the uniform diagnostic standard and data processing method.Results The asthenia of both the heart and the spleen(56 cases, 10 8%) was one of the primary syndrome of EII.Conclusions This study showed that main symptoms in differential diagnosis of asthenia of both the heart and the spleen might be sleeplessness, poor appetite, excessive dream, dizziness and weakness of limb.
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0.05).Expression of CD25and CD69on CD8 + T cells was significantly higher in patients(16.30%?5.97%and32.61%?6.36%)than that in control group(9.21%?2.14%and10.03%?2.35%)(P
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Detection of epidermal cellular DNA content by flow cytometry was performed in normal skin of 14 individuals, both lesiona] and nonlesional areas of 36 patients with psoriasis vulgaris. The results showed that the proliferative activity of epidermal cells was increased successively in normal skins, nonlesional and lesional skin of patients. The differences were significant (P O.01). In combination with the clinical features and histopathological changes and the comparison of 6 patients before and after treatment, the clinical significance of above changes has been discussed.
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Objective To investigate the differentiation of dendritic cells from monocytes of the peripheral blood in patients with psoriasis. Methods Flow cytometry was used to analyze the phenotype of monocyte derived dendritic cells (MoDC). The capacity of MoDC to stimulate the proliferation of T lymphocytes was evaluated by allogeneic mixed lymphocyte reaction. Results Monocytes in the peripheral blood of patients with psoriasis could differentiate into dendritic cells. Expression of CD40, CD80, CD86 and HLA-DR by MoDC was significantly increased in patients with psoriasis compared with that in normal controls (P