原发性胆汁性肝硬化患者Siglec-1蛋白的表达及其临床意义

目的检测Siglec-1(CD169)在原发性胆汁性肝硬化(PBC)患者外周血单核细胞上的蛋白表达,并探讨其与原发性胆汁性肝硬化发生发展的作用及临床意义。方法流式细胞术检测35例原发性胆汁性肝硬化患者及35例健康对照者、35例肝炎后肝硬化对照者外周血CD14CD169双阳性细胞的表达率;生化常规测定所有入选者血清生化指标水平。结果流式细胞术检测结果显示:原发性胆汁性肝硬化组外周血CD14CD169双阳性率为(13.0±2.2)%,显著高于健康对照组(1.0±0.2)%,及肝炎后肝硬化对照组(4.1±0.4)%,差异具有统计学意义(P<0.01)。并且Siglec-1表达与GGT(r=0.44,P<0.01)和ALP水平(r=0.33,P<0.05)密切相关。结论原发性胆汁性肝硬化患者单核细胞表面siglec-1蛋白表达显著增高,说明原发性胆汁性肝硬化患者外周血单核细胞已经发生巨噬细胞化,单核巨噬细胞介导的免疫炎症反应在原发性胆汁性肝硬化发生发展过程中起重要作用。     

蜂胶对成纤维细胞、单核巨噬细胞系基质金属蛋白酶MMP-9表达的抑制作用

目的探讨蜂胶对糖尿病伤口愈合的作用及其机制。方法采用荧光电泳法,检测不同葡萄糖浓度（5、25mmol／L）和不同蜂胶浓度（10、50、100、200μg／ml）处理的体外培养的人成纤维细胞及单核巨噬细胞系的基质金属蛋白酶MMP-9蛋白的表达。结果高浓度葡萄糖（25mmol／L）状态下,MMP-9表达增加,但与低浓度（5mmol／L）比较,差异无统计学意义。蜂胶可显著降低MMP-9的表达,并呈剂量依赖性（P〈．05）。结论高糖能增加单核巨噬细胞系及成纤维细胞MMP-9的表达,可能会影响糖尿病患者伤口的愈合。蜂胶可抑制这些细胞MMP-9的表达,有益于细胞基质的积聚,从而有益于糖尿病患者伤口的愈合。     

创伤／烧伤后单核－巨噬细胞变化及调节措施研究进展

严重创伤、大面积深度烧伤可刺激机体外周血单核细胞和组织中的巨噬细胞参与炎症反应和免疫应答,深入探讨单核－巨噬细胞变化的分子机制及拮抗措施对预防伤后并发脓毒症和多器官功能衰竭尤为重要。本文介绍了单核－巨噬细胞在创伤／烧伤后数量、功能变化及相关分子机制与逆转措施,并对其未来的研究前景进行了展望。     

MMP-9、TIMP-1与CD44在骨肉瘤中的表达和临床意义

目的:探讨骨肉瘤组织中黏附分子MMP-9、TIMP-1与CD44的表达情况。方法:免疫组织化学SP测定MMP-9、TIMP-1与CD44在45例骨肉瘤的表达,分析表达水平与浸润、转移之间的关系。结果:MMP-9、TIMP-1及CD44在骨肉瘤中的表达明显高于骨软骨瘤(P<0.05);MMP-9、CD44阳性表达与浸润有关(P<0.05)。结论:MMP-9、CD44是骨肉瘤侵袭的标志。     

类风湿关节炎患者滑膜组织中p-AKT、Bcl-2、MMP-9的表达水平及临床意义

目的 探讨类风湿关节炎(RA)患者滑膜组织中磷酸化蛋白激酶(p-AKT)、B淋巴细胞瘤-2(Bcl-2)、基质金属蛋白酶-9(MMP-9)的表达水平，以及上述指标在RA发生发展中的意义。方法 选取2013-03至2022-03东部战区总医院骨科住院的RA患者15例和同期行膝关节手术的关节损伤患者10例，分别为RA组和对照组。收集两组患者滑膜组织病理标本及临床资料，对滑膜组织进行病理学观察并进行评分；采用免疫组织化学染色技术检测p-AKT、Bcl-2、MMP-9蛋白在滑膜组织中的表达情况，并与RA疾病活动指标之间进行相关性分析。结果 RA组与对照组在年龄、性别方面差异无统计学意义(P>0.05)。RA组患者WBC、单核细胞计数(MONO)、中性粒细胞计数(NEUT)、C-反应蛋白(CPR)、类风湿因子(RF)、抗环瓜氨酸肽抗体(Anti-CCP)明显高于对照组(P<0.05或P<0.01);RA组患者滑膜组织Krenn’s病理评分明显高于对照组(P<0.05或P<0.01)。与对照组比较，RA组滑膜组织中p-AKT、Bcl-2、MMP-9的表达明显高于对照组...     

2型糖尿病颈动脉粥样硬化斑块的初步研究

目的探讨2型糖尿病患者颈动脉粥样硬化斑块的不稳定性及发生机制。方法收集33例符合下列条件的2型糖尿病死亡患者:既往有明确的脑梗死或短暂性脑缺血发作,有相应的病史、体征以及影象学证据。同时选取年龄、性别、血压、血脂等相匹配的18例无糖尿病的死亡病例作为对照。将51例死者的双侧颈内动脉完整剥离后连续取材,常规固定、脱水、石蜡包埋、切片、HE染色,从每例中挑选一块组织行免疫组化CD3、CD68、SMA、MMP-2和MMP-9染色,光镜观察HE及免疫组化染色切片,并采用图像分析仪测定颈动脉粥样硬化斑块的纤维帽厚度以及SMA、MMP-2和MMP-9的阳性面积百分比。结果在已经发生过缺血性脑卒中的2型糖尿病病例中,颈内动脉粥样硬化斑块中不稳定斑块的比例显著高于对照组。2型糖尿病组的不稳定斑块中,CD68、CD3、MMP-2及MMP-9的表达要明显强于稳定斑块,而SMA的表达正好相反。结论 2型糖尿病颈动脉粥样硬化斑块中不稳定斑块增多,这与巨噬细胞和T细胞的浸润以及MMP分泌增多有关。     

人外周血白细胞黏附分子的辐射效应研究

目的 研究外周血白细胞表面黏附分子的表达及其介导的细胞黏附能力变化与辐射剂量的关系。方法 人外周血细胞经不同剂量照射后不同时间用流式细胞仪双色标记分析不同白细胞表面不同黏附分子表达,特异底物包被微孔板法和结晶紫染色分析检测单核细胞对不同底物的黏附能力。结果 单核细胞表面CD11b和粒细胞表面CD29表达下降与辐射剂量间存在良好量效关系,照射后单核细胞对底物β1-整合素和胶原蛋白I的黏附能力改变与辐射剂量存在一定量效关系。结论 外周血白细胞表面黏附分子表达及功能改变展现出的与辐射剂量间的良好关系,为进一步深入研究将其作为评估生物受照剂量指标的可能打下基础。     

脂多糖刺激树突状细胞前后TLR2、3、4基因表达的检测

目的 检测脂多糖作用于树突状细胞前后TLR2、3、4基因的表达情况。方法 从外周血分离单核细胞，加入rhGM-CSF及rhIL-4，培养的第6天，实验组加入脂多糖刺激24h，对照组不加。分别提取细胞总RNA，行半定量RT-PCR，产物进行琼脂糖凝胶电泳分析。结果 单核细胞经GM-CSF及IL-4诱导7d后的DC表达较高水平TLR2、3、4。脂多糖刺激24h后，TLR2、3、4表达下降，TLR4几乎测不到。结论 树突状细胞在受脂多糖刺激前后TLRs的表达有显著变化，推测和它的功能成熟有关。     

宫颈癌患者行荧光定量PCR技术检测外周血MMP-9mRNA的价值探究

目的探讨分析宫颈癌患者采用荧光定量PCR技术检测外周血MMP-9mRNA的临床价值。方法选取2017年4月-2019年4月收治的宫颈癌患者30例及妇科良性肿瘤患者30例作为研究对象,通过实时荧光定量PCR技术检测患者外周血MMP-9mRNA表达情况,分析MMP-9mRNA表达与临床病理相关性。结果研究组外周血MMP-9mRNA阳性检出率80.00%(24/30);对照组外周血MMP-9mRNA阳性检出率46.67%(14/30);研究组外周血MMP-9mRNA阳性检出率相比对照组明显更高(P<0.05);患者年龄、病理类型、肿瘤分期、病理分化、淋巴结转移是影响MMP-9mRNA表达的因素(P<0.05);淋巴结转移、肿瘤分期是导致MMP-9mRNA表达阳性表达的危险因素。结论针对宫颈癌患者行荧光定量PCR技术检测外周血MMP-9mRNA有助于对患者肿瘤微转移情况进行及时监测与干预,以减少患者肿瘤远处转移的发生率并改善患者预后。     

哮喘粒细胞/巨噬细胞集落刺激因子mRNA表达的研究

目的:观察支气管哮喘外周血单个核细胞(PBMC)粒细胞/巨噬细胞集落刺激因子(GM-CSF)mRNA表达水平.方法:分离受试者PBMC,提取总RNA,采用地高辛配基标记的mGM-CSF cDNA探针,斑点印迹杂交检测PBMC GM-CSF mRNA表达的相对含量.结果:哮喘组GM-CSF mRNA表达的相对含量明显高于慢性支气管炎组和正常对照组(P<0.05),慢性支气管炎组与对照组间无显著性差异(P>0.05).结论:哮喘PBMC GM-CSF mRNA表达水平增高,可能在哮喘发病中起作用.     

Expression levels of matrix metalloproteinase (MMP)-9 and its specific inhibitor TIMP-1, in septic and aseptic arthritis of the knee

Purpose

In cases of septic knee arthritis, there is excess of matrix metalloproteinases (MMPs) over tissue inhibitors of metalloproteinases (TIMPs), due to enhanced expression and activation that are induced by bacteria in comparison with rheumatic or degenerative arthritis. The aim of this study was to explore the expression levels of synovial gelatinase MMP-9 and its specific inhibitor TIMP-1 in septic and aseptic arthritis and their potential use as additional aids to clinical investigation.

Methods

Gelatin zymography and western blot analysis were applied in effusions from knees of the patients with septic (SA—10 patients), rheumatic (RA—10 patients) and osteoarthritis (OA—10 patients).

Results

Zymographic analysis revealed that all samples contained latent MMP-2 activity, albeit activated MMP-2 appeared in more of the septic than aseptic effusions. MMP-9 was not detected in osteoarthritic synovial fluid samples. Only trace amounts of MMP-9 activity were detected in 4 of 10 patients with RA, whereas higher MMP-9 levels were evident in all samples from SA (P = 0.0241). In immunoblotting assays, samples from SA showed significantly higher levels of MMP-9 compared with samples from RA (P = 0.0052), confirming zymographic results. Although no significant difference in TIMP-1 levels was observed, the estimated MMP-9/TIMP-1 ratio of septic effusions was significantly higher compared with aseptic ones (P = 0.0029).

Conclusions

The data presented suggest enhanced expression and activation of MMP-9 in septic native knee arthritis compared with aseptic. The presence of high levels of MMP-9 with concomitantly increased MMP-9/TIMP-1 ratio and activated gelatinases in effusions, independent of neutrophilic counts, may be indicative for infection.     

IL-8在强直性脊柱炎活动期的表达与意义

目的 探讨趋化因子IL-8在强直性脊柱炎(AS)活动期的表达及意义。方法 通过cDNA微阵列及RT-PCR技术检测活动期AS患者和对照组[健康志愿者、类风湿关节炎(RA)及膝关节外伤患者]的外周血、关节液中单个核细胞和滑膜细胞IL-8的表达．比较AS患者组与各对照组IL-8表达情况的差异。结果 在活动期AS患者外周血单个核细胞(PBMC)或关节液单个核细胞(SFMC)中IL-8的表达均明显高于健康志愿者PBMC中的表达,在RA患者PBMC中IL-8的表达亦高于健康志愿者PBMC中的表达．关节滑膜细胞IL-8的表达水平在AS患者也明显高于膝关节外伤患者。结论 IL-8在活动期AS中存在高表达,可能是AS的一种重要的炎症介质,介导AS滑膜甚至其他受累组织炎症的发生和发展。     

破骨细胞在强直性脊柱炎滑膜组织中的分化及其在外周关节骨质破坏病理机制中的作用

目的检测破骨细胞在强直性脊柱炎（AS）外周关节滑膜组织中的分化情况,了解破骨细胞的分化与AS患者外周关节骨质破坏病理改变的相关性。方法应用单克隆抗体,通过免疫组织化学及酶组织化学染色的方法检测13例AS、16例类风湿关节炎（RA）、17例骨关节炎（OA）及6例健康对照关节滑膜组织中CD68蛋白表达及TRAP染色阳性滑膜细胞的分布状况,并通过计算机辅助图像分析系统和半定量分析方法确定CD68分子表达水平及TRAP染色阳性细胞在各研究组滑膜组织中的差异性。结果滑膜组织中均有CD68阳性细胞,AS和RA组CD68表达水平较OA组和健康对照组显著增高（P〈0．05）,其阳性细胞主要分布于滑膜衬里层,衬里下层也见少量表达;OA和健康者滑膜中仅有少量CD68阳性细胞分布。TRAP染色阳性细胞位于滑膜组织衬里层、淋巴细胞聚集区及滑膜软骨交界区,其中AS组阳性细胞百分数显著低于RA组,OA及正常对照组阳性细胞少见。RA组阳性细胞百分数与RANKL表达水平呈正相关（r=0．442,P=0．043）。结论炎症关节滑膜组织中表达CD68分子及TRAP染色阳性滑膜细胞数量的增加为破骨细胞的分化、成熟提供了充足的细胞数量基础,破骨细胞数量和活性上调是造成强直性脊柱炎外周关节骨质破坏的重要前提。     

超声检测类风湿关节炎膝关节滑膜改变及其血流的临床应用价值

目的：通过观察类风湿关节炎（rheumatoid arthritis ,RA ）膝关节滑膜病变的超声声像图表现,分析滑膜病变超声检查结果与RA活动度的相关性,探讨超声检查RA膝关节滑膜病变的临床应用价值。方法对38例RA患者患侧膝关节和20例正常对照者双侧膝关节行超声检查,根据DAS28评分结果将RA患者分组,分别对比各组与对照组的滑膜厚度,髌上囊积液;分析RA组超声检测结果与DAS28评分之间的相关性。结果 RA组滑膜厚度较对照组明显增厚,髌上囊积液明显增多（ P ＜0．05）。RA组膝关节滑膜厚度、关节积液及滑膜内血流信号分级与RA患者DAS28评分之间存在一定的相关性（ P ＜0．05）。结论超声检查对RA膝关节滑膜病变具有一定的的诊断价值,与DAS28评分的相关性较好,可以作为评估RA膝关节滑膜病变常规检查方法之一。     

Evaluation of the degree of clinical rheumatoid arthritis activity based on the concentrations of cytokines TNF-alpha, IL-12, IL-15, and IL-18 in serum and synovial fluid

BACKGROUND/AIM: Experimental in vitro and in vivo investigations in a mouse model have proved that TNF-alpha, IL12, IL-15 and IL18 participate in the pathogenesis of erosive inflammatory arthritis. The aim of this research was to determine the clinical significance of cytokines in the evaluation of the activity of rheumatoid arthritis (RA). METHODS: Inside a 4-year period we followed-up 64 patients with RA as newly ocurred or in the phase of worsening. We observed the clinical manifestation of the disease upon which we divided the patients in to 3 groups: the patients with low active RA, patients with moderate active RA, and the patients with wild active RA. The control group (n=25 patients) included the patients with osteoarthrosis (OA), and arthritis of the knee. In the samples of serum of all of the patients the concentratin of cytokines TNF-alpha, IL-12, IL-15, and IL-18 were determined using the immunoenzymatic methods in mice for human interleukines. By comparing the concentrations in 30 patients with the high, 14 patiens with moderate, and 20 patiens with the mild activity of RA it was determined that the patients with the high degree of the disease activity, had significantly high (p < 0.01; p < 0.05) concentrations of the examined cytokines in blood and synovial fluid as compared to the patients with the moderate and mild active disease. There was a relationship (p < 0.01) between the concentrations of cytokines in blood and synovial fluid with the quantity of the Disease Activity Score in 28 joints. CONCLUSIONS: Cytokines concentrations could be good indicators of the degree of the general activity of RA. This research could contribute to the interpretation of insufficiently well known views of the pathogenesis role and significance of citokines in an active disease.     

肿瘤干细胞标记物CD133与基质金属蛋白酶-9在乳腺癌组织中的表达及生物学意义

目的探讨乳腺癌组织中基质金属蛋白酶-9（MMP-9）及肿瘤干细胞标志（CD133）的表达及其与乳腺癌临床病理参数之间的关系。方法采用免疫组织化学方法,检测109例乳腺癌组织中的CD133及MMP-9蛋白的表达并对结果进行分析。结果①109例乳腺癌组织中MMP-9、CD133蛋白的表达率分别是40.4%（44/109）、46.8%（51/109）,随肿瘤直径增大以及肿瘤分期的增加,MMP-9蛋白的阳性表达率也明显增加,差别有统计学意义（P〈0.05）;而乳腺癌组织中CD133＋肿瘤干细胞数量则随病理级别的升高而增加（P〈0.05）。②109例乳腺癌组织中,CD133＋肿瘤干细胞MMP-9的阳性表达随病理分级增加而增加（P〈0.05）。结论低分化乳腺癌组织中,MMP-9蛋白与CD133＋肿瘤干细胞存在密切关系,CD133＋肿瘤干细胞中MMP-9蛋白的表达可能参与乳腺癌恶性演进,或可成为预后预测的指标之一。     

18F-FDG PET imaging of rheumatoid knee synovitis correlates with dynamic magnetic resonance and sonographic assessments as well as with the serum level of metalloproteinase-3

Purpose The aim of this study was to assess rheumatoid arthritis (RA) synovitis with positron emission tomography (PET) and ¹⁸F-fluorodeoxyglucose (¹⁸F-FDG) in comparison with dynamic magnetic resonance imaging (MRI) and ultrasonography (US).Methods Sixteen knees in 16 patients with active RA were assessed with PET, MRI and US at baseline and 4 weeks after initiation of anti-TNF- treatment. All studies were performed within 4 days. Visual and semi-quantitative (standardised uptake value, SUV) analyses of the synovial uptake of FDG were performed. The dynamic enhancement rate and the static enhancement were measured after i.v. gadolinium injection and the synovial thickness was measured in the medial, lateral patellar and suprapatellar recesses by US. Serum levels of C-reactive protein (CRP) and metalloproteinase-3 (MMP-3) were also measured.Results PET was positive in 69% of knees while MRI and US were positive in 69% and 75%. Positivity on one imaging technique was strongly associated with positivity on the other two. PET-positive knees exhibited significantly higher SUVs, higher MRI parameters and greater synovial thickness compared with PET-negative knees, whereas serum CRP and MMP-3 levels were not significantly different. SUVs were significantly correlated with all MRI parameters, with synovial thickness and with serum CRP and MMP-3 levels at baseline. Changes in SUVs after 4 weeks were also correlated with changes in MRI parameters and in serum CRP and MMP-3 levels, but not with changes in synovial thickness.Conclusion ¹⁸F-FDG PET is a unique imaging technique for assessing the metabolic activity of synovitis. The PET findings are correlated with MRI and US assessments of the pannus in RA, as well as with the classical serum parameter of inflammation, CRP, and the synovium-derived parameter, serum MMP-3. Further studies are warranted to establish the place of metabolic imaging of synovitis in RA.     

雌激素受体阴性乳腺癌中神经胶质瘤致病因子1、基质金属蛋白酶-9的表达及其临床意义

 目的 探讨雌激素受体(estrogen receptor ,ER)阴性乳腺癌中的神经胶质瘤致病因子1(glioma-associated oncogene homolog 1,Gli1)及基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)的表达及临床意义。方法 用免疫组化Envision法检测ER阴性乳腺癌组织中Gli1、MMP-9、 CD34的表达情况, 分析Gli1、MMP-9表达与CD34、肿瘤大小、淋巴转移、临床TNM分期、HER-2表达的关系。结果 ER阴性乳腺癌组织中,Gli1阴性(-)、低表达(+)、中表达(++)及高表达(+++)组的CD34分别为48.83±7.53、60.69±10.02、64.68±12.92和75.91±11.30; MMP-9分别为48.15±8.44、61.69±10.71、67.76±14.03、73.11±12.78,组间比较差异均有统计学意义(P<0.05)。ER阴性乳腺癌组织中Gli1、MMP-9表达与肿瘤大小、临床TNM分期呈正相关(P<0.05)。ER阴性乳腺癌组织中Gli1与MMP-9表达存在一定正相关性(r=0.462,P<0.01)。结论 Gli1与MMP-9表达呈正相关性,推测Gli1是ER阴性乳腺癌增殖途径中重要的调控节点之一。     

NMR studies on human,pathologically changed synovial fluids: Role of hypochlorous acid

Recently, it has been reported that hypochlorous acid (HOCl), a special product of neutrophil myeloperoxidase, degrades N-acetyl groups of N-acetylglucosamine, chondroitin sulfate, hyaluronic acid, and minced articular cartilage via a transient product to acetate. This work concerns ¹H NMR investigations of synovia! fluids of patients with rheumatoid arthritis (RA). Synovial fluids of patients with severe forms of this disease are characterized by enhanced ¹H NMR signals for N-acetyl groups (?2.0 ppm) and acetate (1.90 ppm) and the appearance of a broad but less intense signal at 2.35 ppm. It is likely that this signal corresponds to the transient, chlorinated product of degradation of N-acetyl groups by hypochlorous acid. Moreover, ¹H NMR signal intensities of N-acetyl groups and acetate strongly correlate with the myeloperoxidase activities in synovial fluids from patients with rheumatoid arthritis. These results have been confirmed by treatment of native sheep synovial fluid with sodium hypochlorite, resulting in the formation of the same resonances as observed in pathologically changed synovial fluids from humans. Thus, it is concluded that HOCI plays an important role for the cartilage degradation during rheumatoid arthritis.