基质金属蛋白酶及其组织抑制因子与糖尿病足

创面修复是一个复杂而有序的生物学过程,许多重要过程均受细胞因子和蛋白酶的调节。研究表明糖尿病患者足部伤口中基质金属蛋白酶(MMPs)表达增加,其抑制因子基质金属蛋白酶组织抑制因子(TIMPs)表达减少,导致伤口难以愈合。MMPs与TIMPs的比例可能比它们的绝对浓度更重要,是预测伤口愈合的重要指标,与伤口愈合时间呈负相关。局部应用强力霉素、一些含蛋白酶抑制剂的敷料以及促有丝分裂的牛血清提取物等均可通过抑制MMPs促进创面愈合。     

Coagulation inhibitors in severe sepsis: state of the art

OBJECTIVE: To present and discuss the rationale and the results of clinical trials using supplementation with physiologic anticoagulants (Tissue Factor Pathway Inhibitor (TFPI), AntiThrombin (AT), and Protein C (PC) in patients with severe sepsis. RATIONALE: An early activation of the coagulation cascade occurs in severe sepsis. TFPI, AT, and PC are major inhibitors of the coagulation cascade, and additionally modulate inflammatory and vascular reactions. They are consumed or inhibited in the sepsis pathologic process. Therapeutic supplementation with these inhibitors could improve the sepsis-induced organ failures and mortality. CLINICAL RESULTS: Randomized controlled studies were recently completed. No effect on the mortality rate could be documented after treatment with recombinant TFPI. AT concentrates neither improve mortality, but a biological interaction with heparin therapy could have biased the study results. Treatment with recombinant activated PC (alpha-drotrecogin) was associated with a significant reduction in the mortality rate of severely ill patients and received recently the approval from FDA and EC authorities in this indication. An increase in the rate of hemorrhagic adverse effects has been observed with these compounds, justifying a strict observance of contraindications and of patients selection. PROSPECTIVE: Additional studies are needed to give confirmation of the positive effects of activated PC supplementation in less severely ill patients, children and specific clinical situations. The effects of new anticoagulant compounds are currently evaluated in preclinical studies.     

Matrix metalloproteinases and their tissue inhibitors (TIMPs) in Plasmodium falciparum malaria: serum levels of TIMP-1 are associated with disease severity

BACKGROUND: Molecular mechanisms involved in the pathogenesis of severe malaria caused by Plasmodium falciparum are not fully understood. Matrix metalloproteinases (MMPs) are enzymes that proteolytically degrade both the extracellular matrix and nonmatrix substances with various functions in the modulation of immune response. The key inhibitors of MMPs are the tissue inhibitors of metalloproteinases (TIMPs). METHODS: We studied levels of MMP-8, MMP-9, TIMP-1, and TIMP-2 on admission and after 24 h, using an enzyme-linked immunosorbent assay, in serum specimens from 50 Gabonese children with severe malaria, 43 children with uncomplicated malaria, and 27 healthy control children. RESULTS: Serum MMP-8 and TIMP-1 levels were significantly higher in the severe malaria and uncomplicated malaria groups, compared with those in the control group (P < .001). TIMP-1 levels were significantly higher in patients with severe malaria, compared with those in patients with uncomplicated malaria (P < .001). High TIMP-1 levels were significantly correlated with malaria severity, as determined by the simplified multiorgan dysfunction score (Spearman rank-correlation coefficient, 0.55; P < .001). CONCLUSIONS: TIMP-1 is associated with signs and symptoms of severe malaria. MMP-8 levels are elevated in patients with severe or uncomplicated P. falciparum malaria. MMPs and TIMPs may be relevant in the pathogenesis of severe malaria, either as proteolytic enzymes that degrade the extracellular matrix or as effectors and regulators of the immune response.     

Serum MMP-1 and TIMP-1 levels are increased in patients with psoriatic arthritis and their siblings

OBJECTIVE: To determine matrix metalloproteinase-1 (MMP-1) and tissue-inhibitor metalloproteinase-1 (TIMP-1) serum levels in patients with psoriatic arthritis (PsA) and to compare this with their siblings and local blood donor controls. PsA is an interesting condition in which to study metalloproteinases because there are variations in the level of destructiveness, including a significant proportion of cases without destructive change. This is unlike rheumatoid arthritis (RA) which is more uniformly destructive and where MMP-1/TIMP-1 levels are known to be elevated. METHODS: MMP-1 and TIMP-1 serum levels were determined by enzyme-linked immunosorbent assay (ELISA) in (a) index cases with PsA (subtype: RA n = 43, distal interphalangeal disease n = 2, oligoarticular n = 15, spondyloarthropathy n = 9, enthesitis n = 1), (b) siblings with PsA, (c) siblings with psoriasis (Ps), (d) unaffected siblings and (e) local controls. Patients with Ps were divided according to the onset of disease: type I disease, onset before age 40 yr and type II, onset after age 40 yr. RESULTS: MMP-1 and TIMP-1 levels were significantly increased in both the index cases and the group including all siblings compared with the controls (P < 0.0001). There was no statistical difference in MMP-1 or TIMP-1 levels between index cases and their siblings. There was no difference in serum MMP-1 level between the different subtypes (Moll and Wright) of PsA, but there was an increased level of serum TIMP-1 in patients with rheumatoid pattern (P = 0.05). In the index cases there were increased levels of TIMP-1 in type II onset psoriasis (P = 0.03) but no difference in MMP-1 levels. CONCLUSION: MMP-1 and TIMP-1 serum levels are elevated in PsA. This is greatest in RA pattern PsA. These levels were also elevated in unaffected siblings suggesting that genetic factors may be important. TIMP-1 levels were elevated in psoriasis alone, more so in late onset psoriasis, suggesting that the pathological processes of early and late onset psoriasis may be different.     

Plasma levels of the chemokines monocyte chemotactic proteins-1 and -2 are elevated in human sepsis

Because of their effects on monocytes, monocyte chemotactic proteins-1 and -2 (MCP-1 and MCP-2) may participate in the pathophysiology of sepsis. We measured circulating MCP-1 and MCP-2 levels in 42 septic patients having positive local or blood cultures. MCP-1 and MCP-2 levels were elevated in 24 (57%) and 25 (59%) of 42 septic patients, respectively, compared with healthy volunteers. Both patients with gram- positive and gram-negative infections had elevated MCP-1 plasma levels (P = .0001) and P < .0001), respectively; Mann-Whitney-U test), whereas patients with gram-positive infection, but not those with gram-negative infection, had increased MCP-2 plasma levels (P= .0182). No relative differences in MCP-1 and MCP-2 plasma levels were observed between several subgroups of patients (sepsis v septic shock; survivors v nonsurvivors), although levels of MCP-1 were the highest in patients with the more severe forms of sepsis, ie, those with shock or a lethal outcome. Serial observations showed that MCP-1 and MCP-2 plasma levels remained elevated for at least 48 hours. MCP-1 correlated weakly with interleukin-8 and MCP-2, the correlations for which were most pronounced in patients with septic shock. MCP-2 correlated with interleukin-8, and surprisingly, with the complement activation product C3a; these correlations further improved when analyzing patients with septic shock or when applying gram-positive infections. Thus, our results not only show increased MCP-1 and MCP-2 levels in patients with sepsis, but also suggest that the synthesis and release of MCP-1 and MCP-2 in sepsis are differently regulated in part.     

肌红蛋白变化对脓毒症相关慢性重症患者预后的预测价值

目的:探讨肌红蛋白（Mb）变化对脓毒症相关慢性重症（CCI）患者预后的预测价值。方法:选2017年1月至2020年3月中山大学附属第一医院重症医学科一科诊断为脓毒症相关CCI的患者。收集所有患者基线资料,记录入ICU第1天和第14天患者Mb水平,记录28 d内患者生存情况。Kaplan-Meier曲线分析患者28 d累...     

Independent prognostic value of elevated high-sensitivity C-reactive protein in chronic heart failure

Background

The serum concentration of C-reactive protein (CRP) is mildly elevated in patients with chronic congestive heart failure (CHF), but this level falls well within the range found in healthy subjects. Standard clinical assays for CRP lack sensitivity within the low reference range and thus cannot be used effectively for routine clinical risk prediction. Because assays for high-sensitivity CRP (hsCRP) are now available, we can measure hsCRP to determine its predictive value for the prognosis of patients with CHF.

Methods

Serum levels of hsCRP in 108 patients with CHF and left ventricular ejection fraction (LVEF) <50% were examined. Major adverse cardiac events (death, heart transplantation, or hospitalization with worsening heart failure) during a median follow-up period of 403 days were determined.

Results

The concentrations of hsCRP in this study population were significantly increased with the severity of CHF. In a multivariate analysis, LVEF and serum levels of hsCRP were independent significant predictors for adverse outcomes in these patients (hazard ratio, 3.714, P = .024, and hazard ratio, 2.584, P = .047, respectively). However, hsCRP was minimally correlated with LVEF (r = −0.167, P = .084). Further analysis indicated that hsCRP might identify a different high-risk group and could improve risk stratification beyond that of LVEF.

Conclusions

These findings suggest that an elevated level of hsCRP is an independent predictor of prognosis in CHF and can provide additional prognostic information for the risk stratification and treatment in patients with chronic CHF.     

Expression of MMP-9 and TIMP-1 as prognostic markers in gastric carcinoma

BACKGROUND/AIMS: The aim of the present work is to clarify the role of metalloproteinase-9 and its inhibitor in the evolution of gastric cancer after surgical resection. METHODOLOGY: We have studied 44 gastric cancer patients submitted to surgery. There were 13 proximal tumors, 16 located in the middle third and 15 in the distal one. Overall survival was 26% at 6 years. Metalloproteinase-9 and tissue inhibitor of metalloproteinase concentrations were investigated by means of ELISA in frozen samples of tumoral and normal gastric mucosa. RESULTS: Mean concentration of metalloproteinase-9 in tumoral tissue was 42 ng/mg of total protein, and this value was 6.9 times greater than the mean concentration in non-tumoral tissue. Cancer tissue also expressed higher levels of TIMP-1, 7.25 versus 4.39 ng/mg of protein. Higher levels of metalloproteinase expression in tumoral tissue, greater [metalloproteinase in tumor]/[metalloproteinase in non-tumor] ratio and greater [metalloproteinase]/[inhibitor] ratio in tumor cells, were all of them statistically related to a worse prognosis when T1 and T2 tumors were considered. CONCLUSIONS: The expression of metalloproteinase-9 or its inhibitor is related to a more aggressive phenotype of gastric cancer.     

Immunolocalisation studies on six matrix metalloproteinases and their inhibitors, TIMP-1 and TIMP-2, in synovia from patients with osteo- and rheumatoid arthritis.

OBJECTIVE--To assess the likely importance of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in the arthritic process. METHODS--Synovial samples from seven joints with rheumatoid arthritis and three osteoarthritic joints were analysed by indirect immunofluorescence microscopy. Using specific human antisera, we documented the frequencies and distributions of collagenase, stromelysins 1 and 2, matrilysin, gelatinases A and B, TIMP-1, and TIMP-2. RESULTS--Stromelysin 1 was found in all synovia, bound to extracellular matrix, within cells, or both, indicating stromelysin synthesis. Matrilysin was present in only one active inflammatory synovium, and focal synthesis of collagenase and gelatinase A was seen in four synovia. Stromelysin 2 and TIMP-2 were not observed, but TIMP-1 synthesis was seen in five synovia, and in two active synovia the distribution of TIMP-1 positive cells was more widespread than that of MMPs. CONCLUSIONS--The presence of stromelysin 1 in all synovia clearly implicates this enzyme in joint damage. Collagenase, gelatinase A and matrilysin may also have a role in rheumatoid arthritis, but are not significant in osteoarthritis. However, marked regional variations were found in the synthesis of these MMPs, indicating not only that these diseases are episodic but that control of enzyme synthesis is focal. Only TIMP-1 may be considered an inhibitory factor.     

Clinical prognostic markers in patients with severe sepsis: a prospective analysis of 139 consecutive cases

OBJECTIVES: To analyze the clinical characteristics and determine predictive factors of mortality in previously healthy individuals suffering from severe sepsis. METHODS: The study included 139 patients with severe sepsis, admitted to the Department of Medicine over a two years period. Data recorded on admission included demographic information, blood pressure, core temperature, white blood count, hepatic and renal function tests, coagulation factors, blood gases, serum lactic acid levels, simplified acute physiology score (SAPS-II) and Glasgow Coma Scale (GCS). RESULTS: On admission, 62 patients were hypotensive, 52 had signs of diffuse intravascular coagulation (DIC), 72 had renal and 27 hepatic dysfunction. The overall mortality rate was 27.3%. Twenty-nine patients had septic shock on admission with a mortality rate of 62.07%. Hypoxemia, metabolic acidosis and the presence of DIC were more frequent in non-survivors, who also had significantly higher SAPS-II on admission and days 3 and 7. Independent factors associated with mortality were older age, septic shock, DIC and acute renal failure on admission, as well as SAPS-II at all time points and lactic acid levels on day 7. CONCLUSIONS: Septic patients with advanced age, septic shock, renal failure, DIC and metabolic acidosis on admission are at increased risk of mortality. The sustained presence of high SAPS-II and lactacidemia one week after admission are also important risk factors of poor outcome.     

降钙素原及C反应蛋白水平在重症肺炎患者预后中的作用

目的：探讨降钙素原（Procalcitonin，PCT）及C反应蛋白水平（C-reactionprotein，CRP）在重症肺炎患者预后判断及转归中的作用 方法：2012年2月-2014年12月，回顾性分析我科收治重症肺炎患者65例，按照预后情况分为死亡组和存活组，对比分析两组间的临床一般特征、入院治疗前、48h、72h、出院前或死亡前PCT、CRP水平及APACHEⅡ评分 结果：两组患者PCT、CRP水平均较正常值明显升高，其中死亡组PCT 、CRP水平较存活组明显提高（P<0.01）。通过连续监测PCT、CRP水平发现抗生素使用有效时PCT水平明显降低，而CRP水平下降速度较PCT明显降低。血清 PCT 水平与APACHEⅡ评分的相关系数为 0.689，其相关性均高于CRP。PCT、CRP水平的升高与患者死亡具有相关性，OR值( 95% CI)分别为 2.9 (1.2- 7.8 )、7.4( 3.5 ~ 18.7) (P=0.003,0.001 )。结论：PCT、CRP水平在重症肺炎患者预后判断及转归中具有较好的作用，其中PCT水平监测效果优于CRP水平。     

Different effects of angiogenesis inhibitors IFN-alpha and TIMP-1 on lymphangiogenesis

This study was designed to examine the effects of angiogenesis inhibitors IFN-alpha and TIMP-1 on lymphangiogenesis. We cultured lymphatic endothelial (LE) cells from pig thoracic ducts and performed morphological observations using light microscopy, TEM, and confocal microscopy to confirm their lymphatic origin. We tested these cells for growth inhibition by angiogenesis inhibitors IFN-alpha and TIMP-1 using both the scraping line and MTT methods. In addition, we analyzed apoptosis using the Hoechst and Caspase staining methods. Finally, we tested IFN-alpha and TIMP-1 using in vivo inhibitory assays. By morphological observations, all LE cells in vivo and in vitro were found to be of very similar morphology. Both in vitro inhibitory assays of scraping line and MTT showed significant differences for the IFN-alpha treatment (p < 0.01) and no significant difference for TIMP-1. Hoechst and Caspase apoptosis assays demonstrated that IFN-alpha could induce apoptosis of LE cells, and TIMP-1 had little effect. IFN-alpha and TIMP-1 inhibitory in vivo assays showed a lack of healing following IFN-alpha treatment compared to control and TIMP-1 treatment. In summary, these different angiogenesis inhibitors have different effects on lymphangiogenesis. IFN-alpha inhibits proliferation and migration of LE cells in a dose-dependent fashion and induces apoptosis of LE cells while TIMP-1 has no significant inhibitory effects on proliferation, migration, or inducing apoptosis.     

Circulating matrix metalloproteinases 1, 2, 9 and their inhibitors TIMP-1 and TIMP-2 as serum markers of liver fibrosis in patients with chronic hepatitis C: comparison with PIIINP and hyaluronic acid

OBJECTIVES: Histological examination of liver biopsy is currently required in the management of patients with chronic hepatitis C. Our aim was to evaluate the diagnostic utility of a panel of circulating markers in detecting the stage of fibrosis. METHODS: One hundred and ninety four-patients who had undergone a percutaneous liver biopsy before antiviral treatment, and 194 age- and sex-matched healthy subjects were studied. Serum levels of hyaluronate, procollagen type III N-terminal peptide (PIIINP), matrix metalloproteinases (MMP)-1, MMP-2, MMP-9 and their tissue inhibitors of metalloproteinases (TIMP)-1 and TIMP-2 were determined by RIA and ELISA. Histological lesions were staged according to the METAVIR score. RESULTS: Hyaluronate, PIIINP, TIMP-1, and TIMP-2 serum levels were significantly higher in patients than in controls. Six markers were significantly correlated with fibrosis: MMP-2 (r = 0.28; p < 0.01), TIMP-1 (r = 0.42; p < 0.001), HA (r = 0.50; p < 0.001), PIIINP (r = 0.62; p < 0.0001), MMP-1 (r = -0.32; p < 0.01), and MMP-9 (r = -0.22; p < 0.05). By multivariate analysis, only PIIINP and MMP-1 were independently associated with fibrosis, and were combined using the equation of the logistic regression model. Using receiver-operating characteristics analysis, the area under the curve of the score to discriminate mild (FO/F1) from significant fibrosis (F2/F3/F4) was 0.82, with a sensitivity of 60% for a specificity of 92%. CONCLUSION: Our results suggest that combining two serum markers reflecting fibrogenesis (PIIINP) and fibrolysis (MMP-1) may provide a useful tool for evaluating liver fibrosis.     

Elderly patients with non-cardiac admissions and elevated high-sensitivity troponin: the prognostic value of renal function

BACKGROUND High-sensitivity cardiac troponin(hs-cTn) levels are frequently elevated in elderly patients presenting to the emergency department for non-cardiac events. However, most studies on the role of elevated hs-cTn in elderly populations have investigated the prognostic value of hs-cTn in patients with a specific diagnosis or have assessed the relationship between hs-cTn and comorbidities.AIM To investigate the in-hospital prognosis of consecutive elderly patients admitted to the Internal Medicine Department with acute non-cardiac events and increased hs-cTnI levels.METHODS In this retrospective study, we selected patients who were aged ≥ 65 years and admitted to the Internal Medicine Department of our hospital between January 2019 and December 2019 for non-cardiac reasons. Eligible patients were those who had hs-cTnI concentrations ≥ 100 ng/L. We investigated the independent predictors of in-hospital mortality by multivariable logistic regression analysis.RESULTS One hundred and forty-six patients(59% female) were selected with an age range from 65 to 100(mean ± SD: 85.4 ± 7.61) years. The median hs-cTnI value was 284.2 ng/L. For 72(49%) patients the diagnosis of hospitalization was an infectious disease. The overall in-hospital mortality was 32%(47 patients). Individuals who died did not have higher hs-cTnI levels compared with those who were discharged alive(median: 314.8 vs 282.5 ng/L; P = 0.565). There was no difference in mortality in patients with infectious vs non-infectious disease(29% vs 35%). Multivariable analysis showed that age(OR 1.062 per 1 year increase, 95%CI: 1.000-1.127; P = 0.048) and creatinine levels(OR 2.065 per 1 mg/dL increase, 95%CI: 1.383-3.085; P 0.001) were the only independent predictors of death. Mortality was 49% in patients with eGFR 30 mL/min/1.73 m2.CONCLUSION Myocardial injury is a malignant condition in elderly patients admitted to the hospital for non-cardiac reasons. The presence of severe renal impairment is a marker of extremely high in-hospital mortality.     

急性白血病患者WT1基因与LRP基因关系的研究

目的：探讨急性白血病患者ＷＴ１基因与ＬＲＰ基因表达的关系。方法：应用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）法检测７３例急性白血病患者及２３例正常人的ＷＴ１及ＬＲＰ基因的表达。结果：７３急性白血病患者中４１例ＷＴ１基因表达阳性,２５例ＬＲＰ基因表态阳性;２３例正常人中ＷＴ１基因表达均阴性,仅１例ＬＲＰ基因表达阳性。ＷＴ１基因表达阳性患者的完全缓解（ＣＲ）率低于表达阴性者（分别为５３．７％和８４．４％）     

Serum levels of adhesion molecules ICAM-1 and VCAM-1 and tissue inhibitor of metalloproteinases, TIMP-1, are elevated in patients with autoimmune thyroid disorders: Relevance to vascular inflammation

Serum levels of ICAM-1 (Inter Cellular Adhesion Molecule-1), VCAM-1 (Vascular cell Adhesion Molecule-1-I), TIMP-1 (tissue inhibitor of metalloproteinases 1) and MMP-9 (Metalloproteinase 9) are well established markers of inflammation. The physiopathological link between inflammation, atherosclerosis and autoimmunity is well demonstrated. However, serum levels of these biomarkers in patients with autoimmune-mediated dysthyroidism, including their evolution after improvement of the thyroid disorder have not been assessed. So, we evaluated the circulating levels of these markers in autoimmune and in non-autoimmune-mediated dysthyroid patients, and their evolution after treatment of thyroid disease.We conducted a prospective study to evaluate these markers before and after treatment in hyperthyroid patients (n = 33; 28 patients with autoimmune disease), hypothyroid patients (n = 38; 33 patients with autoimmune disease) and euthyroid subjects (n = 33). At baseline, serum levels of ICAM-1, VCAM-1 and TIMP-1 were significantly elevated in patients with hyperthyroidism as compared to euthyroid and hypothyroid patients (respectively p = 0.0005 and p < 0.0001). In multivariate analysis, the differences remained significant for VCAM-1 and TIMP-1. Median levels of ICAM-1, VCAM-1 and TIMP-1 were significantly higher in patients with autoimmune-mediated dysthyroidism compared to euthyroid patients (respectively p < 0.0001 and p = 0.002). In hyperthyroid patients, ICAM-1, VCAM-1 and TIMP-1 concentrations fell significantly after they had become euthyroid (respectively p = 0.0006; p < 0.0001 and p = 0.0009), although VCAM-1 values remained higher than those observed in the control group (p = 0.005).We found that autoimmune-mediated dysthyroidism were associated with increased peripheral blood concentrations of VCAM-1, ICAM-1 and TIMP-1. Whether these biological abnormalities translate into increase intima remodelling and atherosclerosis remains to be studied.     

Long-term prognostic value of elevated heart rate one year after heart transplantation

Background

Elevated heart rate (HR) is associated with adverse cardiovascular outcome in the general population and in patients with cardiovascular disease. Elevated HR due to graft denervation is often found in heart transplantation (HTx) patients; the effect on graft survival and vasculopathy is unclear. Thus, the aim of this study was to evaluate the role of elevated HR at 12 months post-HTx and its power to predict HTx long-term outcome.

Methods

We evaluated retrospectively a prospective database of 312 patients undergoing HTx at two centers. HR was registered at 12 months post-HTx. The median HR was used as a cutoff point. Cox regression analysis was performed with variables known to be clinically relevant to mortality and those selected from the univariate analysis.

Results

During a mean follow-up of 5.5 ± 2.8 years there were 58 deaths (19%). Patients with a HR ≥ 90 bpm (median HR) at 12 months had an increased risk for all-cause mortality (Hazard Ratio = 2.4, 95% CI 1.2 to 4.5, p = 0.009) and mortality related to coronary allograft vasculopathy (CAV) (Hazard Ratio = 3.0, 95% CI 1.25–7.14, p = 0.01). Multivariate analysis showed that a HR ≥ 90 bpm independently predicted mortality (HR 3.2, 95% CI 1.4–7.1, p = 0.004).

Conclusions

Elevated HR measured at 12 months after HTx is an independent predictor of all-cause mortality in HTx recipients. A HR ≥ 90 bpm identifies a group of patients at high risk of death and CAV-related mortality at mid- to long-term.