Benquan Wu

Tumor-infiltrating lymphocytes predict response to chemotherapy in patients with advance non-small cell lung cancer

Accumulating preclinical evidence suggests that anticancer immune responses contribute to the success of chemotherapy. The predictive significance of tumor-infiltrating lymphocytes (TILs) for response to neoadjuvant chemotherapy in non-small cell lung cancer (NSCLC) remains unknown. The aim of this study was to investigate the prognostic and predictive value of TIL subtypes in patients with advanced NSCLC treated with platinum-based chemotherapy. In total, 159 patients with stage III and IV NSCLC were retrospectively enrolled. The prevalence of CD3+, CD4+, CD8+ and Foxp3+ TILs was assessed by immunohistochemistry in tumor tissue obtained before chemotherapy. The density of TILs subgroups was treated as dichotomous variables using the median values as cutoff. Survival curves were estimated by the Kaplan–Meier method, and differences in overall survival between groups were determined using the Log-rank test. Prognostic effects of TIL subsets density were evaluated by Cox regression analysis. The presence of CD3+, CD4+, CD8+, and FOXP3+ TILs was not correlated with any clinicopathological features. Neither the prevalence of TILs nor combined analysis displayed obvious prognostic performances for overall survival in Cox regression model. Instead, higher FOXP3+/CD8+ ratio in tumor sites was an independent factor for poor response to platinum-based chemotherapy in overall cohort. These findings suggest that immunological CD8+ and FOXP3+Tregs cell infiltrate within tumor environment is predictive of response to platinum-based neoadjuvant chemotherapy in advanced NSCLC patients. The understanding of the clinical relevance of the microenvironmental immunological milieu might provide an important clue for the design of novel strategies in cancer immunotherapy.

Predictive value of MMP‐7 expression for response to chemotherapy and survival in patients with non‐small cell lung cancer

Fundamental studies have suggested that matrix metalloproteinases‐7 (MMP‐7) expression is associated with chemoresistance and constitutes a prognostic factor in several solid tumors. The present study assessed the prognostic and predictive value of MMP‐7 in tumors of patients with advanced non‐small cell lung cancer (NSCLC) treated with platinum‐based chemotherapy. In total, 159 patients with stage III and IV NSCLC were retrospectively enrolled. Immunohistochemistry was performed to evaluate the expression of MMP‐7, apoptosis‐related proteins Bcl‐2, Bax, Fas and FasL and the Ki‐67 proliferation marker. The TUNEL (terminal deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate nick‐end labeling) method was performed to investigate tumor apoptosis. Ninety carcinomas (56.6%) were identified as high expression of MMP‐7. Overexpression of MMP‐7 was more frequent in adenocarcinomas than in squamous cell carcinomas (P = 0.032). The expression of MMP‐7 was positively related with Ki‐67 index and Bcl‐2, but not apoptosis index. MMP‐7 status was correlated inversely with response to chemotherapy in overall patients (response rates, 20.0% and 35.8%, for patients with high‐MMP‐7 and low‐MMP‐7 tumors, respectively, P = 0.036), especially in adenocarcinoma (P = 0.021), but not in patients with squamous cell carcinomas (P = 0.373). The overall survival was significantly lower in NSCLC patients with high MMP‐7 than in those with low MMP‐7 (P < 0.001). A Cox regression analyses also demonstrated MMP‐7 status to be a significant prognostic factor (hazard ratio, 5.49; P = 0.001). These findings suggest that the expression level of MMP‐7 in tumor cells is predictive of response to chemotherapy and outcome in patients with advanced NSCLC receiving platinum‐based chemotherapy. (Cancer Sci 2008; 99: 2185–2192)

Factors associated with the outcome of life-threatening necrotizing pneumonia due to community-acquired Staphylococcus aureus in adult and adolescent patients.

Background: Although community-acquired Staphylococcus aureus pneumonia with highly virulent Panton-Valentine leukocidin (PVL)-positive strains, a severe disease with significant lethality, is rare, especially in adult and adolescent patients, recent reports highlight that these infections are on the rise. Objectives: To describe the demographic and clinical features of reported cases of life-threatening community-acquired S. aureus pneumonia with usually PVL-positive strains in adult and adolescent patients, to evaluate the variables related to death, and to select a more appropriate antimicrobial treatment for this potentially deadly disease. Methods: We summarized all of the 92 reported cases and our case. The effect of 5 variables on mortality was measured using logistic regression. Results:S. aureus community-acquired pneumonia (CAP) with usually PVL-positive strains is a severe disease with significant lethality, i.e. 42.9%; a short duration of the time from the onset of symptoms to death, i.e. 5.5 ± 10.1 days, and prolonged hospital admissions, i.e. 33.2 ± 29.5 days. Seventy-three cases have been tested for the gene for PVL, and 71 strains have been found to carry the PVL gene. Logistic regression analysis showed that leucopenia (p = 0.002), influenza-like symptoms or laboratory-confirmed influenza (p = 0.011), and hemoptysis (p = 0.024) were the factors associated with death. Antibiotic therapies inhibiting toxin production were associated with an improved outcome in these cases (p = 0.007). Conclusions: Physicians should pay special attention to those patients who acquired severe CAP during influenza season and have flu-like symptoms, hemoptysis, and leucopenia, and they should consider S. aureus more frequently among the possible pathogens of severe CAP. Empiric therapy for severe CAP with this distinct clinical picture should include coverage for S. aureus. Targeted treatment with antimicrobials inhibiting toxin production appears to be a more appropriate selection.

Inhibitory effects of simvastatin on staphylococcus aureus lipoteichoic acid-induced inflammation in human alveolar macrophages

Staphylococcus aureus(S. aureus) is the most common bacterium in sepsis and pneumonia involving gram-positive bacteria. Lipoteichoic acid (LTA) is a cell wall component of gram-positive bacteria. It is a potent inducer of inflammatory mediators in human dendritic cells, human pulmonary epithelial cells, and murine macrophages. However, the effect of LTA on human alveolar macrophages (AMs) which are the major effector cells in host defense against respiratory tract infections has hardly been studied. Statins have anti-inflammatory, immunomodulatory, antioxidative, anticoagulant, and antibacterial activities. These effects may be contributed to reduce the markers of systemic inflammation. Emerging retrospective studies have demonstrated that statin use decreased the mortality of pneumonia. However, the precise mechanisms responsible for these effects are unclear. The purpose of this study is to define the role of S. aureus LTA in human AMs and the effects of simvastatin (SV) on LTA-stimulated human AMs. The results showed that LTA induced tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), IL-8 mRNA expression, and suppressed IL-10 mRNA expression in human AMs. Simultaneously, LTA induced human AMs apoptosis. These effects were parallel with the up-regulation of the expression of NF-κB-P65 protein in the LTA-stimulated human AMs. The above effects of LTA on human AMs were inhibited significantly by SV. These data indicate that S. aureus LTA induces potent pro-inflammatory and pro-apoptotic effects on human AMs and statins exert anti-inflammatory effects by mediating inhibition of NF-κB activation and cytokine mRNA expression in human AMs. These results may explain, in part, the mechanisms responsible for favorable effects of statins on pneumonia.

STAT1 regulates MD-2 expression in monocytes of sepsis via miR-30a.

Sepsis is a major cause of morbidity and mortality in critically ill patients. MD-2 is a 25-kDa lipopolysaccharide (LPS)-binding protein that forms a heterodimer with TLR42, but its regulation in sepsis is not clear. This study aims to investigate the molecular mechanism of regulation of MD-2. Inflammation cytokines in monocytes were analyzed by real-time RT-PCR and ELISA, and it was found that IL-10 was elevated significantly in the monocytes with LPS treatment. And then, when the cells were treated with IL-10, STAT1 was activated in the monocytes using Western blotting. It was also found that STAT1 could enhance MD-2 expression on transcriptional and posttranscriptional levels. Finally, miR-30a was predicted to the molecule that may regulate STAT1 expression. It was verified that STAT1 was a new target gene of miR-30a. miR-30a could inhibit IL-10-induced cytokine release by targeting STAT1–MD-2 in monocytes. In conclusion, this study for the first time demonstrated that miR-30a inhibits MD-2 expression by targeting of STAT1 in human monocytes.

Matrilysin inhibits proliferation and modulates sensitivity of lung cancer cells to FasL-mediated apoptosis.

Aims and background Matrix metalloproteinase (MMP) family member MMP-7 (matrilysin) cleaves various cell-surface proteins to alter their effector functions in addition to a broad substrate specificity against extracellular matrix components. Matrilysin expression is closely associated with the advanced clinicopathological stages and unfavorable prognosis. The current study tried to describe the comprehensive impacts of matrilysin on proliferation, and focused on its influence on the susceptibility to FasL-induced apoptosis in A549 lung adenocarcinoma cell line. We also detected the expressions of apoptosis-relative genes to further clarify the underlying mechanisms. Study design The viability of A549 cells was determined by MTT and the apoptosis was assessed by Hoechst 33342 staining and Annexin V-FITC/PI apoptosis kit. The expressions of apoptosis-relative genes were evaluated by flow cytometry, ELISA, and real-time quantitative RT-PCR, respectively. Results Overall, matrilysin exhibited the inhibition of cell growth in a dose- and time-dependent manner by arresting in G0/G1 phase of the cell cycle and inducing apoptosis on A549 cells. Although it directly promoted apoptosis at high concentrations, a certain range of matrilysin might protect tumor cells from FasL-mediated death. The underlying mechanism may be due to the imbalance in the susceptibility of surface membrane-bound Fas receptor and ligand to proteolysis activity of matrilysin. Conclusion Our data indicated matrilysin may be multiple, multifarious, and multifaceted functions contributing to early tumor growth. A therapeutic key might be to modulate activity and function of matrilysin under diverse pathological conditions, but not completely eliminate the expression or function.

Protective effects of caffeoylxanthiazonoside isolated from fruits of Xanthium strumarium on sepsis mice

Abstract Context: The fruit of Xanthium strumarium L. (Asteraceae) has been used for the treatment of various inflammatory diseases. Objective: This study investigates the protective effect of caffeoylxanthiazonoside (CYXD) isolated from fruits of X. strumarium on sepsis mice in vitro and in vivo. Materials and methods: Cecal ligation and puncture (CLP) operation was used to establish the sepsis mice model, and sham mice were also performed. CYXD was administered by intraperitoneal injection (10, 20, and 40 mg/kg/d), then the survival rate was measured in 96 h. Additionally, sepsis mice were induced by injection LPS (2 mg/kg); CYXD was administered by intraperitoneal injection (10, 20, and 40 mg/kg/d), then mice were sacrificed, and serum levels of TNF-α and IL-6 were determined by ELISA assay. Furthermore, the ability of CYXD to neutralize LPS was measured by using the LAL test, and expressions of TNF-α, IL-6 were determined by using real-time fluorogenic PCR. Results: Results indicated that CYXD significantly elevated survival rates of sepsis mice induced by CLP (p < 0.05) with survival rates of 35%, 45%, and 65%. Furthermore, the LPS level was decreased obviously by CYXD (1, 2, and 4 mg/L) (p < 0.05). Additionally, CYXD (10, 20, and 40 mg/kg) can not only significantly decrease TNF-α and IL-6 levels induced by LPS in mices serum (p < 0.05), but also inhibit mRNA expressions of TNF-α and IL-6 induced by LPS in RAW 264.7 cells at doses of 20, 40, and 80 μg/mL (p < 0.05). Conclusion: Our study demonstrated that CYXD has significant protective effects on sepsis mice.

Regulatory effects of miR-155 and miR-146a on repolarization and inflammatory cytokine secretion in human alveolar macrophages in vitro

Abstract Macrophages play an important role in inflammatory responses; however, miRNA-mediated repolarization of macrophages is essential for fulfilling this function. To clarify a series of changes at the RNA level in alveolar macrophages under normal and inflammatory conditions, bronchial alveolar lavage liquid (BALF) was collected from healthy volunteers or patients with pneumonia. This approach, which differs from that used in previously, provides more accurate information about the states of macrophages in different lung microenvironments. In this study, the density plots of macrophage subtypes (M1 and M2) in the BALF of healthy volunteers differed from that of the patients with pneumonia. The M2 subtype dominated in healthy volunteers and was rapidly repolarized to M1 in response to miRNA-mediated gene regulation. Differential miRNA expression in the two macrophage subtypes revealed lower expression of miR-155 and MIR-146a in patients with pneumonia compared with healthy volunteers; this may be related to inflammation and the use of anti-inflammatory drugs. We also found increased TNF-α and IL-6 expression at the RNA level, while macrophage galactose-type C-type lectin 1 (MGL-1) expression decreased with downregulation of miR-155 and miR-146a expression. These results indicate that the gene regulation mediated by miR-155 and miR-146a contributes to human alveolar macrophage phenotype repolarization, thus leading to an early switch from pro-inflammatory to anti-inflammatory cytokine production.

Comparing therapeutic and adverse effects of moxifloxacin and levofloxacin in treatment of community acquired pneumonia: A meta-analysis of randomized controlled trials

The aim of this study was to compare more conclusively the efficacy and safety of moxifloxacin, a new respiratory fluoroquinolone antibiotic with levofloxacin therapy which has been reported to possess good efficacy for community acquired pneumonia (CAP) in hospitalized elderly patient. The entire patients were mild to moderate by CAP. All the trials administered intravenously moxifloxacin (400 mg daily) and intravenously levofloxacin (200500 mg daily) for 7-14 days. Clinical response during therapy was between days 3 and 5 after the start of therapy. A meta-analysis of randomized controlled trials (RCTs, identified in CNKI, PubMeD, Embase, Google) were performed. Eight RCTs involving 1310 patients were included in metaanalysis. Two reviewers independently extracted data from published trials that compared fluoroquinolones. A meta-analysis was performed with the clinical outcomes of moxifloxacin and levofloxacin. Moxifloxacin monotherapy was associated with better therapeutic effect [(OR=1.63, 95%CI 1.01-2.63) and similar adverse effect (OR=1.12, 95%CI 0.81-1.55)] compared with levofloxacin therapy for CAP. Moxifloxacin has better therapeutic effects with comparable adverse events compared to levofloxacin.