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1.
Li JJ  Wei W  Shi HZ  Li YX  Mo WN 《中华医学杂志(英文版)》2011,124(24):4160-4165
Background  Interleukin 16 (IL-16) can be detected by ELISA in pleural effusion (PE) and its concentration is higher than in serum. This study investigated the cellular sources of IL-16 in PE.
Methods  The samples of PE were collected from 34 patients who were newly diagnosed having PE in the pleural cavity. We performed cell culture to purify the pleural mesothelial cells (PMC), Wright staining to count the purity and immunocytochemical stain to identify the cultured cells. The intracellular IL-16 expression was detected by flow cytometry (FCM). The different cells in PE were first separated by magnetic cell sorting (MCAS) then the separated cells were cultured in RPMI1640 with 10% fetal calf serum (FCS). We extracted the supernatant and detected IL-16 concentration by ELISA. The IL-16 protein was detected by immunohistochemistry and double immunofluorescence staining.
Results  The percentages of cells which secreted IL-16 were: CD3+CD8 cells ((74.27±15.56)%, n=34); CD3+CD8+ cells ((69.86±18.55)%, n=34); CD19+ cells ((45.30±18.77)%, n=15); CD14+ cells ((16.91±16.69)%, n=15); and PMC ((2.05±1.85)%, n=7). The concentrations of IL-16 in the supernatant from cultured cells were: CD4+ cells ((102.50±42.51) ng/L, n=5); CD8+ cells ((92.58±18.34) ng/L, n=5); CD19+ cells ((79.85±5.62) ng/L, n=5); CD14+ cells ((58.51±25.38) ng/L, n=5); and PMC ((18.14±8.37) ng/L, n=5). In lymphocytes, monocytes/macrophages and PMC, we could observe the cells that expressed IL-16 protein. In paraffin-embedded sections, we also could observe by immunohistochemistry the CD4+IL-16+ cells, CD8+IL-16+ cells, CD19+IL-16+ cells, and CD14+IL-16+ cells.
Conclusions  IL-16 in PE is mainly secreted by T lymphocytes, including CD3+CD8 cells and CD3+CD8+ cells. CD19+ cells and CD14+ cells can also secrete IL-16, but the percentage of PMC that can secrete IL-16 is very low.
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2.
穿龙薯蓣中抗血栓活性成分研究   总被引:2,自引:0,他引:2  
目的 为进一步寻找穿龙薯蓣Dioseorea nipponica抗血栓活性成分,对其甾体皂苷类化学成分进行了系统的分离。方法 采用正相、反相等色谱分离手段进行分离纯化,通过物理化学方法,根据MS、NMR谱数据,对得到的化合物进行结构鉴定。结果 共分离得到9个化合物,分别鉴定为薯蓣皂苷元(1)、薯蓣皂苷元-3-O-β-D-葡萄糖苷(2)、薯蓣皂苷元-3-O-[α-L-鼠李糖基(1→2)]-β-D-葡萄糖苷(3)、薯蓣皂苷元-3-O-[α-L-鼠李糖基(1→4)]-β-D-葡萄糖苷(4)、薯蓣皂苷(5)、纤细皂苷(6)、伪原薯蓣皂苷(7)、26-O-β-D-葡萄糖基-3β,26-二醇-25(R)-Δ5,20(22)-二烯-呋甾-3-O-[α-L-鼠李糖基(1→2)]-O-β-D-葡萄糖苷(8)、26-O-β-D-葡萄糖基-3β,26-二醇-25(R)-Δ5,20(22)-二烯-呋甾-3-O-[α-L-鼠李糖基(1→4)]-O-β-D-葡萄糖苷(9)。结论 化合物3479为首次从该植物中分离得到。大鼠静脉旁路血栓形成试验表明,化合物25显示出一定的抗血栓活性。  相似文献   

3.
目的 研究玉簪Hosta plantaginea的抗炎活性部位及其化学成分。方法 萃取法制备不同极性部位,醋酸致小鼠腹腔毛细血管通透性法、棉球致小鼠肉芽肿法筛选抗炎活性部位;硅胶柱色谱等方法对活性部位进行分离纯化,根据理化性质和波谱数据进行结构鉴定。结果 醋酸乙酯部位能明显抑制小鼠腹腔毛细血管通透性增高以及棉球肉芽肿;从醋酸乙酯部位分离并鉴定了10个化合物,分别为二十二烷醇(1)、β-谷甾醇(2)、豆甾醇(3)、(25R)-2α,3β-二羟基-5α-螺旋甾烷-9(11)-烯-12-酮(4)、胡萝卜苷(5)、(25R)-2α,3β-二羟基-5α-螺旋甾烷-9(11)-烯-12-酮3-O-{O-β-D-吡喃葡萄糖基-(1→2)-O-[β-D-吡喃木糖基-(1→3)]-O-β-D-吡喃葡萄糖基-(1→4)-β-D-吡喃半乳糖苷}(6)、山柰酚3-O-(2″-O-β-D-吡喃葡萄糖基)-β-D-芸香糖苷(7)、山柰酚3-O-β-D-芸香糖苷-7-O-β-D-吡喃葡萄糖苷(8)、(25R)-2α,3β,12β-三羟基-5α-螺旋甾烷3-O-[O-α-L-吡喃鼠李糖基- (1→2)-β-D-吡喃半乳糖苷](9)、(25R)-2α,3β-二羟基-5α-螺旋甾烷3-O-[O-β-D-吡喃葡萄糖基-(1→4)-β-D-吡喃半乳糖苷](10)。结论 醋酸乙酯部位为玉簪抗炎的活性部位;化合物110均为首次从玉簪中分离得到,其中化合物10为新的天然产物。  相似文献   

4.
Background  Patients with xanthogranulomatous cholecystitis sometimes exhibit imaging and intraoperative findings that are similar to those of advanced gallbladder cancer, thus these patients are easily misdiagnosed. The present study aimed to investigate the characteristics of xanthogranulomatous cholecystitis masquerading as gallbladder cancer that could potentially aid in the correct diagnosis of this condition.
Methods  The clinical, serological, radiological and operative features of twelve patients with obviously wall-thickening or mass-forming xanthogranulomatous cholecystitis were retrospectively analyzed. Additionally, the patient preoperative features were compared to those of 36 patients with advanced gallbladder cancers.
Results  Twelve patients with xanthogranulomatous cholecystitis exhibited one to three episodes of acute cholecystitis within 0.5 to 7 months prior to admission to the hospital. Five of these patients exhibited concomitant choledocholithiasis, whereas no concomitant choledocholithiasis was identified in patients with advanced gallbladder cancer. The incidence of abdominal pain (χ2=6.588, P=0.010), acute cholecystitis (χ2=29.176, P=0.000), acute cholangitis (χ2=6.349, P=0.012), choledocholithiasis (χ2=16.744, P=0.000), carcinoembryonic antigen test (P=0.007), CA125 (P=0.001), and diffuse gallbladder wall thickening (χ2=6.031, P=0.014), continued mucosal line (χ2=15.745, P=0.000), homogeneous enhancement of mucosal line (χ2=19.947, P=0.000), submucosal hypoattenuated nodules or band (χ2=18.607, P=0.000) in computed tomography demonstrated statistically significant differences between cases of xanthogranulomatous cholecystitis and gallbladder cancer. Furthermore, all the twelve patients with xanthogranulomatous cholecystitis exhibited at least one positive computed tomography imaging feature aside from past acute cholecystitis episode, and no patient with advanced gallbladder cancer simultaneously exhibited past acute cholecystitis episode and at least one positive computed tomography imaging feature.
Conclusions  The accurate preoperative diagnosis of xanthogranulomatous cholecystitis includes an integrated review of past acute cholecystitis episode, choledocholithiasis, and positive computed tomography imaging features. Besides, we present an algorithm for intraoperative diagnosis.
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5.
Background  Copernicus optical coherence tomography (SOCT) is a new, ultra high-speed and high-resolution instrument available for clinical evaluation of optic nerve. The purpose of the study was to compare the agreements between SOCT and Heidelberg retinal tomography (HRT).
Methods  A total of 44 healthy normal volunteers were recruited in this study. One eye in each subject was selected randomly. Agreement between SOCT and HRT-3 in measuring optic disc area was assessed using Bland-Altman plots. Relationships between measurements of optic nerve head parameter obtained by SOCT and HRT-3 were assessed by Pearson correlation.
Results  There was no significant difference in the average cup area (0.306 vs. 0.355 mm, P=0.766), cup volume (0.158  vs. 0.130 mm, P=0.106) and cup/disc ration (0.394 vs. 0.349 mm, P=0.576) measured by the two instruments. However, other optic disc parameters from SOCT were significantly lower compared with HRT-3. The Bland-Altman plot revealed good agreement of cup area and cup volume measured by SOCT and HRT-3. Bad agreement of disc area, rim area, rim volume and cup/disc ratio were found between SOCT and HRT-3. The highest correlations between the two instruments were observed for cup area (r2=0.783, P=0.000) and cup/disc ratio (r2=0.669, P=0.000), whereas the lowest correlation was observed for disc area (r2=0.100, P=0.037), rim area (r2=0.275, P=0.000), cup volume (r2=0.005, P=0.391) and rim volume (r2=0.021, P=0.346).
Conclusions  There were poor agreements between SOCT and HRT-3 for measurement of optic nerve parameters except cup area and cup volume. Measurement results of the two instruments are not interchangeable.
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6.
Donor MHC gene to mitigate rejection of transplantation in recipient mice   总被引:1,自引:0,他引:1  
Li T  Yan J  Tan JL  Lü YP  Hou SC  Li ST  Xu Q  Tong XH  Ding J  Zhang ZT  Li H 《中华医学杂志(英文版)》2011,124(24):4279-4285
Background  Donor organ rejection continues to be a significant problem for patients receiving transplants. We therefore tested whether transferring a donor’s major histocompatibility complex (MHC) gene to the recipient would mitigate the rejection of transplanted hearts in mice. 
Methods  H-2Kk gene from donor mice was amplified using nested polymerase chain reaction (PCR) and ligated into a mammalian expression vector, which was then transfected into thymus ground mass cells collected from the recipients. Clones stably expressing the transgene were then injected into the recipients’ thymus visualized using ultrasound. Control mice were administered cells previously transfected with empty vector. Following heart transplantation, cardiac activity was monitored electrocardiographically. Recipient thymus cells were tested for MHC antigenicity using flow cytometry and spleen cells were subjected to mixed lymphocyte culture tests. Finally, the transplanted hearts were sectioned, stained and examined under light microscopy.
Results  Southern analysis following nested PCR revealed clear expression of H-2Kk gene. Following transplantation, electrocardiosignals were detectable highly significantly longer in recipients administered thymal cells expressing donor H-2Kk than in those receiving control cells. Flow cytometric analysis using an anti-H-2Kk antibody confirmed its expression in H-2Kk treated recipients but not in control mice. Mixed lymphocyte cultures containing H-2Kk treated cells showed significantly less proliferation than those containing control cells. Hearts from control mice showed substantially greater lymphocyte infiltration than those from H-2Kk treated mice and large areas of necrosis.
Conclusion  Rejection of transplanted hearts can be mitigated substantially by introducing the donor’s MHC into the recipient.
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7.
Background  Previous studies indicate that CD43 plays a role in regulating the adhesion of lymphocytes, cell mutation and activation, however, little is known about its effect on systemic lupus erythematousus (SLE). This study was designed to explore the clinical significance of CD43 in SLE patients.
Methods  We used microarray and real-time PCR to detect the mRNA and protein expression of magnetic bead sorted T cells and B cells from peripheral blood mononuclear cells (PBMCs) of SLE patients, and analyzed the relationship between CD43 and the clinical indexes.
Results  Both microarray and real-time PCR results showed that CD43 mRNA was significantly decreased in PBMCs of SLE patients compared with healthy controls (P <0.001). There were no significant differences between lupus nephritis and non-lupus nephritis patients, and neuropsychiatric and non-neuropsychiatric patients. CD43 mRNA expression was significantly reduced in T cells but not in B-cells in SLE patients compared to healthy controls (P <0.01). Compared with healthy controls, the percentage of CD43+ cells in the PBMCs of SLE was significantly decreased (P=0.004), and the CD43 fluorescence intensity in CD3+/CD43+ cells and CD19+/CD43+ cells was also significantly weaker than in healthy controls (P=0.039 and 0.003). There was no significant difference in the percentage of CD3+/CD43+ cells, CD19+/CD43+ cells between the two groups. The CD43 fluorescence intensity in CD3+/CD43+ cells was inversely correlated with the levels of IgG and IgM (r= 0.8 and –0.6).
Conclusions  Compared to healthy controls, both CD43 mRNA and protein expressions were reduced in T cells from patients with SLE, and were inversely correlated with IgG.
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8.
Background  Regulatory T cell populations, particularly CD4+CD25+ T regulatory cells, have been implicated in the persistence of hepatitis B virus (HBV) infection. However, no clear relationship has been established between the frequency of CD4+CD25+ T regulatory cells in the peripheral blood and either the disease phases in the natural history of chronic HBV infection or in the response to interferon-α therapy.
Methods  In the present study, three different common markers of CD4+CD25+ T regulatory cells were used to determine the numbers of T regulatory cells in healthy controls and in patients with chronic HBV infection.
Results  No significant difference was found when samples were gated for CD25hi and CD25+FoxP3+ T cells. A significant correlation was found between the number of CD4+ Treg cells that gated with CD25+FoxP3+ and CD25+CD127low/- in healthy controls and in patients with chronic hepatitis B (CHB) (r=0.67, 0.59; P <0.01). The percentages of Treg cells were (8.56±2.01)% in asymptomatic carriers (Asc), (8.74±3.04)% in inactive HBsAg carriers, (10.7±2.93)% in CHB and (7.42±1.28)% in healthy controls (F=11.1, P <0.001). The percentage of Treg cells in patients with CHB was higher than in asymptomatic HBV patients, inactive HBsAg carriers, or healthy controls (P <0.01). The proportion of CD4+CD25+CD127low/- T cells in patients who responded to interferon-α was (11.9±3.3)%, (9.1±2.4)% and (9.0±2.9)% at baseline, week 12 and week 24 after treatment, respectively (Z=2.42, P <0.05; Z=2.67, P <0.01).
Conclusions  These results suggest that the proportion of the CD4+CD25+ regulatory T cells might be affected by the application of different markers in process to detect T regulatory cells. The frequency of Treg cells was increased in patients with CHB, which might be associated with the disease activity of these patients and contribute to prevention of extensive liver damage. A decline in Treg cells at week 12 of treatment might be associated with a better response to treatment with interferon-α.
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9.
Background  It has been reported that the nicotinic acetylcholine receptor subunit α4 gene (CHRNA4) might be associated with smoking behaviors in the previous studies. Up to now, there are few reports on the relationship between CHRNA4 and smoking initiation. In this study, we tried to explore the role of two polymorphisms in CHRNA4 (rs1044396 and rs1044397) in smoking initiation and nicotine dependence in Chinese male smokers.
Methods  Nine hundred and sixty-six Chinese male lifetime nonsmokers and smokers were assessed by the Fagerström test for nicotine dependence (FTND), smoking quantity (SQ) and the heaviness of smoking index (HSI). All subjects were divided into four groups based on their tobacco use history and the FTND scores. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to find two polymorphisms of CHRNA4 in these subjects.
Results  The χ2 test showed that rs1044396 was significantly associated with smoking initiation (χ2=4.65, P=0.031), while both rs1044396 and rs1044397 were significantly associated with nicotine dependence (χ2=5.42, P=0.020; χ2=7.58, P=0.005). Furthermore, the T-G (3.9%) haplotype of rs1044396-rs1044397 showed significant association with smoking initiation (χ2=6.30, P=0.012) and the C-G haplotype (58.9%) remained positive association with nicotine dependence (χ2=8.64, P=0.003) after Bonferroni correction. The C-G haplotype also significantly increased the HSI (P=0.002) and FTND scores (P=0.001) after Bonferroni correction.
Conclusion  These findings suggest that CHRNA4 may be associated with smoking initiation and the C-G haplotype of rs1044396-rs1044397 might increase the vulnerability to nicotine dependence in Chinese male smokers.
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10.
Background  Allergen-specific immunotherapy can induce immune tolerance to specific allergens by regulating immune status of individuals. However, its clinical application is limited due to individual differences in efficacy among patients and un-confirmed safety. 1,25 Dihydroxyvitamin D3 (1,25(OH)2D3) has been shown to be involved in a variety of physiological processes, including immune response regulation. In the present study we explored the role of 1,25(OH)2D3 pretreatment for immunotherapy.
Methods  Seventy-five BALB/c mice were randomly divided into five groups (15 mice per group). The mouse allergic asthma model was established by intra-peritoneal injection of ovalbumin (OVA, 10 μg) and aluminium hydroxide (2 mg) as an adjuvant. Intra-peritoneal injection of 50 ng of 1,25(OH)2D3 served as a pretreatment, subcutaneous injection of OVA (100 μg) as an immunotherapy, and 1% OVA inhalation as a challenge. Histopathological analysis was performed on four mice per group. The number of cells and their classification in bronchoalvolar lavage (BAL) fluid were assayed. Levels of serum OVA-specific immunoglobulin E (sIgE) and IFN-γ, IL-4, IL-5 and IL-10 in BAL fluid were measured by ELISA.
Results  After 1,25(OH)2D3 pretreatment, immunotherapy could significantly inhibit the infiltration of inflammatory cells into lung tissues and BAL fluid of mice with allergic asthma when compared with un-treated animals (eosinophils: (7.46±1.34)×104/ml vs. (13.41±1.67)×104/ml, P <0.05). In addition, levels of IL-4 ((36.91±7.87) pg/ml vs. (43.70±6.42) pg/ml, P >0.05) and IL-5 ((41.97±7.93) pg/ml vs. (60.14±8.35) pg/ml, P <0.05) in BAL fluid and serum sIgE ((0.42±0.05) vs. (0.75±0.06) OD units, P <0.05) were profoundly reduced. However, the IL-10 level in BAL fluid was significantly increased ((67.74±6.57) pg/ml vs. (44.62±8.81) pg/ml, P <0.05).
Conclusions  These results indicated that 1,25(OH)2D3 pretreatment enhanced the inhibitory effects of immunotherapy on allergic airway inflammation. In the treatment of allergic diseases, 1,25(OH)2D3 pretreatment may be beneficial for improving the efficacy of immunotherapy. 
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