Tissue inhibitor of metalloproteinase‐1 levels in plasma from tumour arteries and veins of patients with rectal cancer

Objective. Tissue inhibitor of metalloproteinase‐1 (TIMP‐1) plays a major role in the regulation of tissue growth, including cancer growth. The TIMP‐1 protein can be determined in plasma, and increased plasma levels of TIMP‐1 are associated with a poor prognosis of colorectal cancer patients. The aim of the present study was to evaluate whether tumour tissue release of the TIMP‐1 protein contributes to the increased plasma levels of TIMP‐1 observed in patients with colorectal cancer. Material and methods. Preoperative blood samples from a peripheral vein and intraoperative blood samples from a tumour artery, a tumour vein and from a peripheral vein were drawn from 24 patients undergoing elective, intended curative surgery for primary rectal cancer. TIMP‐1 levels were determined concurrently in plasma from all samples using a validated ELISA method. Counts of white blood cells and platelets were also carried out. Results. No significant differences between plasma TIMP‐1 levels could be demonstrated in any compartment. In particular, there was no significant difference in TIMP‐1 levels in plasma from tumour arteries and tumour veins. However, there was a significant decrease in neutrophil cell counts from tumour arteries to tumour veins (p<0.001). Conclusions. The present results do not support the current hypothesis that tumour cells contribute substantially to increased plasma TIMP‐1 levels observed in patients with colorectal cancer.     

Assessment of the clinical significance of antigenic and functional levels of α1-proteinase inhibitor (α1-Pi) in infiltrating ductal breast carcinomas

ObjectivesTo determine the clinical significance of α1-proteinase inhibitor (α1-Pi) in infiltrating ductal breast carcinoma patients.Design and methodsSerum levels of α1-Pi, tryptic specific inhibitory capacity and α1-Pi circulating immune complexes were determined using radial immunodiffusion, BAPNA assays and ELISA, respectively. 2-DE-MS and immunohistochemistry were performed to examine α1-Pi protein expression.ResultsA decreased serum level of α1-Pi was found among breast cancer patients in comparison to controls. In addition, we found a significantly decreased mean level of α1-Pi in the node metastatic group when compared to node negative patients. However, the functional activity of the inhibitor did not decrease proportionately. Through 2-DE analyses, a differential expression of α1-Pi isoforms according to tumor stage and node metastatic development was found.ConclusionsBoth α1-Pi levels and specific activity could be a source of complementary clinical information and may provide useful information for a better understanding of the mechanisms of metastasis.     

Seasonal variation and stability of matrix metalloproteinase-9 activity and tissue inhibitor of matrix metalloproteinase-1 with storage at -80°C

Objective

To determine whether active matrix metalloproteinase (MMP)-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 displayed seasonal variation and were stable in storage.

Methods

Plasma active MMP-9 and TIMP-1 were measured at three time-points in 163 individuals.

Result

There was no evidence for seasonal variation or declining levels for up to three years of storage at − 80 °C.

Conclusion

Active MMP-9 and TIMP-1 appear to be stable seasonally, and in storage for at least three years.     

Determination of elastase inhibitory activity of α1-proteinase inhibitor and bronchial antileukoprotease: Different results using insoluble elastin or synthetic low molecular weight substrates

We have determined the effect of two elastase-specific synthetic low molecular weight substrates, L-pyroglutamylprolylvaline-paranitroanilide and succinyl-trialanylparanitroanilide, together with insoluble elastin-fluorescein, on the determination of the neutrophil elastase (NE) inhibitory capacity of purified α₁-proteinase inhibitor (α₁-PI) and bronchial antileukoprotease (ALP). In addition, the inhibitory capacities of mixtures of α₁-proteinase inhibitor, antileukoprotease and α₂-macroglobulin prepared in ratios similar to that in lung secretions were determined. Purified inhibitors, alone or in combinations, inhibited about 1 mole neutrophil elastase per mole inhibitor when assessed using synthetic substrates. However, when elastin-fluorescein was used in the assay system, the purified inhibitors showed an inhibitory capacity that was 40–85% of the value obtained using synthetic substrates. Even less inhibition was observed when mixtures of inhibitors were assessed using elastin-fluorescein (23–44% of the value for synthetic substrates). Our data indicate that results of elastase inhibitor activity measurements depend on the type of substrate which has been used.     

C1 inhibitor: just a serine protease inhibitor? New and old considerations on therapeutic applications of C1 inhibitor

C1 inhibitor is a potent anti-inflammatory protein as it is the major inhibitor of proteases of the contact and the complement systems. C1-inhibitor administration is an effective therapy in the treatment of patients with hereditary angioedema (HAE) who are genetically deficient in C1 inhibitor. Owing to its ability to modulate the contact and complement systems and the convincing safety profile, plasma-derived C1 inhibitor is an attractive therapeutic protein to treat inflammatory diseases other than HAE. In the present review we give an overview of the biology of C1 inhibitor and its use in HAE. Furthermore, we discuss C1 inhibitor as an experimental therapy in diseases such as sepsis and myocardial infarction.     

Changes of plasma endothelin, thromboxame B2and 6-keto-PGF1αin patients with coronary heart disease after PTCA

Objective To explore the regularity of changes of plasma endothelin (ET), throm-boxame B2(TXB2) and 6-keto-prostaglandin F1α(6-keto-PGF1α) and restenosis after percutaneous transluminal angioplasty (PTCA) in patients with coronary heart disease. Methods Radioimmunoas-say was applied to measuring plasma levels of ET, TXB2and 6-keto-PGF1αat 0,30 min, 1 day and 3 days after PTCA in 41 patients with coronary heart disease. Results The level of ET in the patients with coronary heart disease was significantly decreased in 30 minutes after PTCA (P＜0.05), but no significant difference was observed in 1 day and 3 days after PTCA (P>0.05). The level of plasma TXB2has no statistical difference after PTCA in 30minutes, 1 day and 3 days (P>0.05). The level of 6-keto-prostaglandin F1α(6-keto-PGF1α) of the patients with coronary heart disease was significantly declined in 30 minutes after PTCA (P<0.05) ,but no significant difference was observed in 1 day and 3 days after PTCA (P>0.05). Conclusion PTCA may lead to fluctuation of plasma levels of ET, TXB2and 6-keto-PGF1αThe clarification of changing regularity of these vasoactive substances contrib-utes to prevention of acute artery occlusion or restenosis after PTCA.     

Role of lactoferrin and C-Reactive protein in ascites of liver cirrhosis for diagnosis of spontaneous bacterial peritonitis北大核心

Objective This study was to investigate the role of lactoferrin and C-reactive protein (CRP) assay in ascitic fluid for diagnosis of spontaneous bacterial peritonitis (SBP) in the patients with decompensated liver cirrhosis. Methods Ascites was collected from the inpatients with decompensated liver cirrhosis before and after treatment from May to December 2011 for analysis of polymorphonuclear cells (PMN) and bacterial culture. The level of lactoferrin and C-reactive protein in the ascites were determined. Results A total of 117 ascites samples were collected from 66 patients. Twenty-six patients met the criteria of SBP with PMN ≥ 250 × 106/L in ascites, were assigned to SBP group. Of these patients, 11 presented with fever 37.3’C to 38C, and 11 patients had elevated peripheral blood white cell count > 10 × 109/L. Eleven patients had neutrophil cell percentage > 0.75. Only 8 patients in this group had positive bacterial culture. Another 12 patients met the criteria of suspected SBP, and assigned to suspected SBP group. The remaining 28 patients did not satisfy the criteria of SBP, and assigned to non-SBP group. The pretreatment lactoferrin level was (68. 46 ± 611.70) ng/mL and (98. 28 ± 56. 81) ng/mL in SBP and non-SBP group, respectively. The pretreatment CRP levll 8. 13 mg/L, respectively. The cu-off value of lactoferrin for diagnosis of SBP was 233 ng/mL with sensitivity 96. 2% and specificity 97. 5%. The cu-off value of CRP for diagnosis of SBP was 4. 390 mg/L with sensitivity 92. 3% and specificity 92. 5%. However, lactoferrin combined with CRP had a sensitivity of 99. 70% and specificity of 90. 18% for diagnosis of SBP. Conclusions Lactoferrin and CRP levels in the ascites of patients with liver cirrhosis are useful for diagnosis of SBP with high specific-ty and sensitivity. © by Editorial Department of Chinese Journal of Infection and Chemotherapy.     

Effect of l-NAME, an inhibitor of nitric oxide synthesis, on plasma levels of IL-6, IL-8, TNFα and nitrite/nitrate in human septic shock

Objectives: We tested the effects of N^G-nitro-L-arginine methyl ester (l-NAME), an inhibitor of nitric oxide (NO) synthesis, on plasma levels of interleukin (IL) IL-6, IL-8, tumor necrosis factor-alpha (TNFα) and nitrite/nitrate (NO ₂ ^? /NO ₃ ^? ) in patients with severe septic shock. Design: Prospective clinical study. Setting: Surgical intensive care unit at a university hospital. Patients: 11 consecutive patients with severe septic shock. Interventions: Standard hemodynamic measurements were made and blood samples taken at intervals before, during, and after a 12-h infusion of l-NAME 1 mg · kg^?1 ·h^?1 for determination of plasma IL-6, IL-8, TNFα and NO ₂ ^? /NO ₃ ^? concentration. Measurements and results: Patients with sepsis had increased plasma levels of IL-6, IL-8, TNFα, and NO ₂ ^? /NO ₃ ^? (p<0.05). Plasma levels of IL-6, IL-8, and NO ₂ ^? /NO ₃ ^? were negatively correlated with systemic vascular resistance (r=?0.62, r=?0.65, and r=?0.78, respectively, all p<0.05). Continuous infusion of l-NAME increased mean arterial pressure and systemic vascular resistance, with a concomitant reduction in cardiac output (all p<0.01). No significant changes were seen in levels of plasma IL-6, IL-8, and NO ₂ ^? /NO ₃ ^? during the 24-h observation period. Plasma levels of TNFα were significantly reduced during l-NAME infusion compared to baseline (p<0.05). Conclusions: NO plays a role in the cardiovascular derangements of human septic shock. Inhibition of NO synthesis with l-NAME does not promote excessive cytokine release in patients with severe sepsis.     

Errors in fracture diagnoses in the emergency department – characteristics of patients and diurnal variation

Background

Evaluation of the circumstances related to errors in diagnosis of fractures at an Emergency Department may suggest ways to reduce the incidence of such errors.

Methods

Retrospective analysis of all cases during a two year period (2002–2004) where a fracture had been overlooked or an injury had been erroneously diagnosed as a fracture (n = 61). 100 random selected patients with correctly diagnosed fractures served as control group.

Results

In the two year period 5879 patients visited the ED with injuries. 1% of all visits to the ED resulted in an error in fracture diagnosis and 3.1% of all fractures were not diagnosed at the initial visit to the ED. 86% of such errors had consequences for treatment. No patient characteristics could be identified as risk factors for a misdiagnosis of a fracture. There was a peak in errors in fracture diagnoses between 8 pm and 2 am (47% against 20% in controls, p < 0.005).

Conclusion

A considerable number of fractures were not correctly diagnosed at the initial ED visit. There was a diurnal variation in the rate of misdiagnosis of fractures with a significant peak from 8 pm to 2 am. Where there was an error in fracture diagnosis, the patients did not appear to have a characteristic profile as regarding e.g. age, sex or capability to communicate with the ED staff. Increased consultancy service in radiology may reduce the frequency of errors in diagnosis, particularly in the evenings between 8 pm and 2 am.     

Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/MS method

ObjectivesCamostat mesilate is a drug that is being repurposed for new applications such as that against COVID-19 and prostate cancer. This induces a need for the development of an analytical method for the quantification of camostat and its metabolites in plasma samples. Camostat is, however, very unstable in whole blood and plasma due to its two ester bonds. The molecule is readily hydrolysed by esterases to 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA) and further to 4-guanidinobenzoic acid (GBA). For reliable quantification of camostat, a technique is required that can instantly inhibit esterases when blood samples are collected.Design and methodsAn ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-ESI-MS/MS) using stable isotopically labelled analogues as internal standards was developed and validated. Different esterase inhibitors were tested for their ability to stop the hydrolysis of camostat ester bonds.ResultsBoth diisopropylfluorophosphate (DFP) and paraoxon were discovered as efficient inhibitors of camostat metabolism at 10 mM concentrations. No significant changes in camostat and GBPA concentrations were observed in fluoride-citrate-DFP/paraoxon-preserved plasma after 24 h of storage at room temperature or 4 months of storage at −20 °C and −80 °C. The lower limits of quantification were 0.1 ng/mL for camostat and GBPA and 0.2 ng/mL for GBA. The mean true extraction recoveries were greater than 90%. The relative intra-laboratory reproducibility standard deviations were at a maximum of 8% at concentrations of 1–800 ng/mL. The trueness expressed as the relative bias of the test results was within ±3% at concentrations of 1–800 ng/mL.ConclusionsA methodology was developed that preserves camostat and GBPA in plasma samples and provides accurate and sensitive quantification of camostat, GBPA and GBA by UHPLC-MS/MS.     

Research updates on pharmacokinetic/pharmacodynamic variation of meropenem in different physiological and pathological states北大核心CSCD

[No abstract available]     

Experimental study on expression of TNF-αand ICAM-1in cerebral ischemia-reperfusion injury

Researchesinrecentyearsfoundthatabnormalleukoaggluti－nationandleukocytosisintheareaofcerebralischemia－reperfusionareimportantfactorscausingtheischemia－reperfusioninjuryandsupposedthatinflammatorymechanismplayedaroleinthereperfu－sioninjury．Advancedresearchesprovedthattumornecrosisfactor－α（TNF－α）andintercellularadhesionmolecule－１（ICAM－１）playedanimportantroleinadhesion，agglutination，leukocytosisfromthebloodvesselsandtherealizationofcytotoxicfunctionoftheleukocytes．Theobjecti…     

Indole-3-carbinol enhances the resolution of rat liver fibrosis and stimulates hepatic stellate cell apoptosis by blocking the inhibitor of κB kinase α/inhibitor of κB-α/nuclear factor-κB pathway

Hepatic stellate cells (HSC) play a pivotal role in liver fibrosis, and the clearance of activated HSC by apoptosis is associated with the resolution of liver fibrosis. The development of strategies that promote this process in a selective way is therefore important. We evaluated the effects of indole-3-carbinol (I3C), a nutritional component derived from vegetables from the Brassica family, on liver fibrosis and HSC apoptosis. The in vivo therapeutic effects of I3C were monitored in three rat models of liver fibrosis induced by porcine serum, bile duct ligation, or multiple hepatotoxic factors, and its proapoptotic effect and molecular mechanism were studied in vitro in HSC-T6, a rat HSC line. The results showed that I3C treatment significantly reduced the number of activated HSC in the livers of rats with liver fibrosis. In histopathology, I3C reduced hepatocyte degeneration and necrosis, accelerated collagen degradation, and promoted the reversal of liver fibrosis. I3C prescribed to HSC-T6 resulted in morphologic alterations typical of apoptosis and DNA cleavage to a nucleosomal ladder. Moreover, I3C significantly increased the HSC-T6 apoptosis rate and the expression ratio of Bax to Bcl-2. High-throughput protein array analysis indicated that the tumor necrosis factor-α/nuclear factor-κB (NF-κB) signal pathway participated in I3C-induced HSC-T6 apoptosis. Western blot and electrophoretic mobility-shift assay confirmed that I3C inhibited the phosphorylation of inhibitor of κB kinase α and inhibitor of κB-α and NF-κB DNA binding activity. In conclusion, I3C could promote the reverse process of liver fibrosis in vivo and induce apoptosis of activated HSC in vitro, which indicates the use of I3C as a potential therapeutic agent in liver fibrosis treatment.     

Effect of irradiation on the quantity and activity of P65,TNF-αand IL-1 in rat wounds

Ourpresentexperimentsindicatedthatatcellularlevels,totalbodyirradiationwithlargedosehasbeenshowntoinhibitinflam-mation,decreasecytokineproduction,suppresscellproliferationandinduceapoptoticcelldeath.Withrespecttomolecularmechanisms,littleisknownabouttheeffectsofionizingradiationonhealingintheearlystage.NF-Bplayimportantrolesinin-flammationprocess.NF-Bactivation,inturn,leadstotheex-pressionofmanygenesinvolvedinimmunologicalandinflammatoryresponses,includinggenesthatencodecytokines,adhesionmol…     

Activity of selected enzymes in erythrocytes and level of plasma antioxidants in response to single whole‐body cryostimulation in humans

The influence of extremely low temperatures on the human body and physiological reactions is not fully understood. The aim of this research was to estimate the influence of a single exposure to cryogenic temperature (?130°C), without subsequent kinesiotherapy, on the activity of the most crucial antioxidant enzymes in erythrocytes: superoxide dismutase (SOD), catalase (CAT), glutathione reductase (R‐GSSG), glutathione peroxidase (GPx) and glutathione transferase (T‐GSH). In the plasma, the concentrations of glutathione, uric acid, albumins and extra‐erythrocyte haemoglobin as components of the non‐enzymatic antioxidant system were evaluated. The subjects were 10 healthy young men. Blood was sampled in the morning on the day of cryostimulation, 30?min after cryostimulation and the next morning. The enzymatic response of the antioxidant defence to the influence of the extremely low temperature resulted in an immediate, significant, increase in GPx and R‐GSSG activities, but a decrease in CAT and T‐GSH activities. We observed an increase in the concentrations of all the examined non‐enzymatic antioxidants, especially extra‐erythrocyte haemoglobin and uric acid, which had both increased further the day after cryostimulation. The results indicate that a single stimulation with cryogenic temperatures results in oxidative stress in a healthy body, but that the level of stress is not very high. It seems that in this case the most significant role in the antioxidant mechanisms is played by peroxidase.     

The changes of plasma 8-iso-prostaglandin F2αand serum C-reactive protein levels in patients with obstructive sleep apnea-hypopnea syndrome

Objective To investigate the plasma 8-iso-prostaglandin F2α(8-iso-PGF2α and the serum C-re-active protein(CRP) levels in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) with and without hypertension(OSAHS + HT),and to explore the changes of pathophysiology in patients with OSAHS and the patho-genesis of OSAHS + HT. Methods All observed subjects were divided into 3 groups: control group (n=20),OS-AHS group(n=19),OSAHS + HT group (n=21). Plasma 8-iso-PGF2αand serum CRP concentrations levels were measured by ELISA and were compared. Results The plasma 8-iso-PGF2αand serum CRP levels,were higher in OSAHS patients than those in control subjects [(11.08±3.26)μg/L vs (7.49±2.10)μg/L,P<0.01;(1.75±0.82) mg/L vs (0.52±0.26 ) mg/L,P<0.01],and were higher in OSAHS + HT group than those in control group [14.84±3.43)μG/L vs(11.08±3.26)μg/L,P<0.01 ;(3.13±1.06)mg/L vs(1.75±0.82)mg/L,P<0.01]. Conclusions Oxidative stress and inflammation in OSAHS patients are increased,which are involved in the devel-opment of OSAHS associated hypertension.     

Intra- and inter-individual variation of BIS-index® and Entropy® during controlled sedation with midazolam/remifentanil and dexmedetomidine/remifentanil in healthy volunteers: an interventional study

Introduction

We studied intra-individual and inter-individual variability of two online sedation monitors, BIS^® and Entropy^®, in volunteers under sedation.

Methods

Ten healthy volunteers were sedated in a stepwise manner with doses of either midazolam and remifentanil or dexmedetomidine and remifentanil. One week later the procedure was repeated with the remaining drug combination. The doses were adjusted to achieve three different sedation levels (Ramsay Scores 2, 3 and 4) and controlled by a computer-driven drug-delivery system to maintain stable plasma concentrations of the drugs. At each level of sedation, BIS^® and Entropy^® (response entropy and state entropy) values were recorded for 20 minutes. Baseline recordings were obtained before the sedative medications were administered.

Results

Both inter-individual and intra-individual variability increased as the sedation level deepened. Entropy^® values showed greater variability than BIS^® values, and the variability was greater during dexmedetomidine/remifentanil sedation than during midazolam/remifentanil sedation.

Conclusions

The large intra-individual and inter-individual variability of BIS^® and Entropy^® values in sedated volunteers makes the determination of sedation levels by processed electroencephalogram (EEG) variables impossible. Reports in the literature which draw conclusions based on processed EEG variables obtained from sedated intensive care unit (ICU) patients may be inaccurate due to this variability.

Trial registration

clinicaltrials.gov Nr. NCT00641563.