Clinicopathological and prognostic significance of myofibrillogenesis regulator-1 protein expression in pancreatic ductal adenocarcinoma.

Myofibrillogenesis regulator-1 (MR-1) expression was detected in different malignancies and is associated with poor prognosis. However, its role in pancreatic ductal adenocarcinoma (PDAC) has not been fully elucidated. Thus, the aim of this study was to investigate the association of MR-1 expression with clinicopathologic features and prognosis in patients with PDAC. Immunohistochemistry was performed to investigate the protein expression of MR-1 and epithelial (E)-cadherin in 87 patients with PDAC. Results showed that MR-1 expression was correlated with histologic grade, tumor stage, and lymph node metastasis (all P <0.05). In addition, MR-1 expression showed a significant inverse correlation with E-cadherin expression (P = 0.002). Furthermore, the variables associated with prognosis were analyzed by Cox's proportional hazards model. Kaplan-Meier analysis was used to plot survival curves according to different expression levels of MR-1. Kaplan-Meier analysis revealed that MR-1 expression was significantly associated with worse disease-free survival (DFS) and overall survival (OS) rates in patients with PDAC (both P <0.001). Finally, multivariate analysis demonstrated that MR-1 expression, together with histologic grade, tumor stage, lymph node metastasis, was an independent prognostic factor for both DFS and OS rates in patients with PDAC. MR-1 overexpression was tightly associated with more aggressive tumor behavior and a poor prognosis, indicating that MR-1 is a valuable molecular biomarker for PDAC progression.

[miR-378 suppresses HBV-related hepatocellular carcinoma tumor growth by directly targeting the insulin-like growth factor 1 receptor].

OBJECTIVE: To investigate the impact and mechanism of microRNA (miR)-378 on hepatocellular carcinoma (HCC) cell growth. METHODS: The human hepatoma cell line HepG2 and its derivative HepG2.2.15 (stably expressing hepatitis B virus (HBV)) were transduced with lentiviruses expressing miR-378 or non-expressing controls (nc-Lv). Effects on cell proliferation were assessed by the MTT assay and on colony-formation efficiency by clonogenic assay. Targets of miR-378 were predicted by bioinformatic analysis and validated by luciferase reporter assay in the human embryonic kidney cell line HEK293. Real-time polymerase chain reaction and western blotting were used to monitor expression of the endogenous targets in miR-378- overexpressing HepG2 and HepG2.2.15 cells. RESULTS: The HepG2 and HepG2.2.15 cells transduced with lentivirus expressing miR-378 showed significantly lower cell proliferation and colony formation than the control cells transduced with nc-Lv (P less than 0.01 and P less than 0.05, respectively). The insulin-like growth factor 1 receptor (IGF1R) was predicted as a potential target of miR-378, and luciferase reporter activity of IGF1R was significantly decreased in the HEK293 cells co-transfected with miR-378 (by 41.8% vs. the control cells, P less than 0.01). Moreover, the miRNA-378-mediated effect was narrowed down to the 3'-untranslated region (UTR) of IGF1R. The miRNA-378-mediated reduction of IGF1R specifically involved its protein expression (P less than 0.01) and not its mRNA expression (P more than 0.05). CONCLUSION: miR-378 may suppress growth characteristics of HBV-related HCC by directly targeting the IGF1R 3'-UTR and inhibiting its protein expression.

Efficacy of ultrasound guided superior laryngeal nerve block on sedation for delayed extubation in maxillofacial surgery with free flap reconstruction.

OBJECTIVE: Superior laryngeal nerve block (SLNB) is a regional anesthesia technique for addressing airway response. However, SLNB on the efficacy of sedation in patients with delayed extubation is unknown, particularly for maxillofacial surgery (MS). The aim of the study was to assess whether ultrasound guided (UG) SLNB reduces the incidence of moderate to severe cough for delayed extubation in MS with free flap reconstruction. METHODS: 60 patients were randomly assigned to the GEA group (control group) and the SLNB group (UG-SLNB postoperatively, study group). During the initial two postoperative hours, the incidence of moderate and severe cough, agitation, and the number of patients requiring rescue propofol and flurbiprofen were recorded. Additionally, the time spent under the target level of sedation, postoperative hemodynamics, and the total does of propofol during the postoperative 24 h were recorded. RESULTS: The data showed the SLNB group had a significantly lower incidence of moderate to severe cough and agitation (p < 0.05), and a longer sedation time (p < 0.05). The number of patients required rescue propofol and flurbiprofen, as well as the hemodynamic changes, were significantly different between the two groups (p < 0.05). CONCLUSION: The use of UG-SLNB is associated with reduced incidence of postoperative cough. Moreover, SLNB can enhance the efficacy of postoperative sedation with need of fewer agents postoperatively. CLINICAL TRIAL REGISTRATION: ChiCTR2000039982.

Metal-organic frameworks constructed from versatile [WS4Cu(x)](x-2) units: micropores in highly interpenetrated systems.

Five metal-organic frameworks (MOFs) formed by [WS(4)Cu(x)](x-2) secondary building units (SBUs) and multi-pyridyl ligands are presented. The [WS(4)Cu(x)](x-2) SBUs function as network vertexes showing various geometries and connectivities. Compound 1 contains one-dimensional channels formed in fourfold interpenetrating diamondoid networks with a hexanuclear [WS(4)Cu(5)](3+) unit as SBU, which shows square-pyramidal geometry and acts as a tetrahedral node. Compound 2 contains brick-wall-like layer also with a hexanuclear [WS(4)Cu(5)](3+) unit as SBU. The [WS(4)Cu(5)](3+) unit in 2 is a new type of [WS(4)Cu(x)](x-2) cluster unit in which the five Cu(+) ions are in one plane with the W atom, forming a planar unit. Compound 3 shows a nanotubular structure with a pentanuclear [WS(4)Cu(4)](2+) unit as SBU, which is saddle-shaped and acts as a tetrahedral node. Compound 4 contains large cages formed between two interpenetrated (10,3)-a networks also with a pentanuclear [WS(4)Cu(4)](2+) unit acting as a triangular node. The [WS(4)Cu(4)](2+) unit in 4 is isomeric to that in 3 and first observed in a MOF. Compound 5 contains zigzag chains with a tetrahedral [WS(4)Cu(3)](+) unit as SBU, which acts as a V-shaped connector. The influence of synthesis conditions including temperature, ligand, anions of Cu(I) salts, and the ratio of [NH(4)](2)WS(4) to Cu(I) salt on the formation of these [WS(4)Cu(x)](x-2)-based MOFs were also studied. Porous MOF 3 is stable upon removal and exchange of the solvent guests, and when accommodating different solvent molecules, it exhibits specific colors depending on the polarity of incorporated solvent, that is, it shows a rare solvatochromic effect and has interesting prospects in sensing applications.

Amentoflavone inhibits colorectal cancer epithelial-mesenchymal transition via the miR-16-5p/HMGA2/ß-catenin pathway.

Background: Amentoflavone is a type of bioflavonoid that exists in many Chinese medicines and has anti-inflammatory, antioxidant, antiviral, and anticancer effects. However, the effect of amentoflavone on epithelial to mesenchymal transition (EMT) in human colorectal cancer (CRC) has not been studied. In this study, we aim to explore the effect of amentoflavone on EMT in CRC. Methods: The effects of long noncoding RNA (lncRNA) miR-16-5p on proliferation, migration, and invasion were determined by in vitro and in vivo experiments. A luciferase reporter assay was carried out to reveal the interaction between miR-16-5p and targeted genes. Reverse transcription polymerase chain reaction (RT-PCR) was used to evaluate the expression of miR-16-5p. A western blot assay was used to detect the expression of targeted genes in CRC cells. Results: The results showed that amentoflavone significantly inhibited CRC migration, invasion, and EMT by increasing miR-16-5p expression. Mechanistically, amentoflavone induced inactivation of the Wnt/ß-catenin pathway via miR-16-5p, directly targeting 3'-UTR of HMGA2 to suppress HMGA2 expression in CRC. Clinically, combined miR-16-5p and HMGA2 levels may serve as a predictor for poor prognosis in patients with CRC. Furthermore, an in vivo PDX model suggested that amentoflavone exhibited antitumor effects in vivo via the miR-16-5p/HMGA2/ß-catenin pathway. Conclusions: This is the first study to show that amentoflavone inhibits CRC EMT via the miR-16/HMGA2/ß-catenin pathway. Amentoflavone may be beneficial in treating CRC patients in the clinic.

Solvatochromic behavior of a nanotubular metal-organic framework for sensing small molecules.

A nanotubular metal-organic framework (MOF), {[(WS(4)Cu(4))I(2)(dptz)(3)]·DMF}(n) (dptz = 3,6-di(pyridin-4-yl)-1,2,4,5-tetrazine, DMF = N,N-dimethylformamide) for sensing small solvent molecules is presented. When accommodating different solvent molecules as guests, the resulting inclusion compounds exhibit different colors depending on the solvent guests, and more interestingly, the band gaps of these solvent-included complexes are in linear correlation with the polarity of the guest solvents. The solvent molecules can be sensed by the changes of UV-vis spectra of the corresponding inclusion compounds, showing a new way of signal transduction as a new kind of sensor. The sensing by such a MOF occurs within the channel-containing material rather than on the external surface.

[Rapamycin-mediated interaction of spleen γδT lymphocytes and lung macrophages in acute lung injury of mice induced by lipopolysaccharide].

OBJECTIVE: To explore the influences of rapamycin (RAPA) upon the cytokine changes of activated spleen γδT lymphocytes and lung tissue macrophages in acute lung injury of mice induced by lipopolysaccharide (LPS). METHODS: A total of 24 healthy male C57BL/6 mice, 6-8 weeks old, were randomly divided into phosphate buffered saline (PBS), LPS, RAPA and LPS + RAPA groups. Acute lung injury was induced by a single intratracheal instillation of LPS in mice. And spleen γδT lymphocytes and lung macrophages were purified by immunomagnetic beads at Day 1. The purified spleen γδT lymphocytes and lung macrophages were adjusted to 10(6) cell/ml. And 24-well plates were used for 4 groups. Each group were further separated with spleen γδT lymphocytes alone, lung tissue macrophages alone and co-culturing. Supernatant fluid was collected after 24 hours. The expressions of IFN-γ and TNF-α were analyzed by ELISA (enzyme-linked immunosorbent assay). And the expressions of mRNA were analyzed by real-time quantitative PCR (polymerase chain reaction). RESULTS: The total cells numbers and lymphocytes numbers of bronchoalveolar lavage fluid were significantly higher in LPS group than those in PBS and LPS + RAPA groups (P < 0.05). And the level of IFN-γ was significantly higher in LPS group than that in PBS, RAPA and LPS + RAPA groups by co-culture (P < 0.05). The level of TNF-α was significantly higher in LPS group than that in RAPA gand LPS + RAPA groups by co-culture (P < 0.05). However, the mRNA of IFN-γ was higher in LPS group than that in PBS and RAPA groups (P < 0.05). CONCLUSION: RAPA inhibits the secretion levels of IFN-γ and TNF-α in spleen γδT lymphocytes and lung tissue macrophages in acute lung injury of mice induced by LPS.

[ γδ T lymphocyte function and the polymorphism of T cell receptor V δ chain in lungs of asthmatic patients].

OBJECTIVE: To observe the function of gamma delta T lymphocytes and the polymorphism of T cell receptor V delta chain in the lungs of asthmatic patients and explore the role of gamma delta T cells in airway inflammation. METHODS: Bronchoalveolar lavage fluid BALF was obtained from 7 asthmatic patients and 7 healthy control individuals. The percentage of gamma delta T cell in BALF was measured by flow cytometry. The gamma delta T cell in BALF was purified by magnetic labeled beads. Proliferous activity was examined by MTT assay. Cytokines secreted by gamma delta T cells in medium was assessed by enzyme-linked immunosorbent assay. Polymorphism of T cell receptor V delta chain was detected by RT-PCR and gene scan analysis. RESULTS: The proportion of gamma delta T cell in the BALF of asthmatic patients [(6.39+/-0.71)%] was significantly higher than that in control subjects [(2.62+/-0.37)%] (P<0.01). The proportion of macrophage in the BALF of asthmatic patients [(81+/-4)] was significantly lower than that in control subjects [(86+/-2)] (P<0.05). The proliferation rate of asthmatic patients [(284.2+/-43.6)%] was significantly higher than that of control subjects [(217.5+/-59.5)%] (P<0.05). Interleukin-4 secreted by gamma delta T cells of asthmatic patients [(18.9+/-3.1) pg/ml)] significantly increased when compared with the control subjects [(14.1+/-3.0) pg/ml] (P<0.05). The polymorphism of T cell receptor V delta chain was not significantly different between these two groups. CONCLUSIONS: The increase of gamma delta T cells in the lung of asthmatic patients further exacerbates Th1/Th2 disturbance and airway inflammation. Antigen recognition by gamma delta T cells is non-specific.

MicroRNA-449b-3p inhibits epithelial-mesenchymal transition by targeting IL-6 and through the JAK2/STAT3 signaling pathway in non-small cell lung cancer.

MicroRNAs (miRs) have vital involvement in the advancement of non-small cell lung cancer (NSCLC); however, the methods of action of miR-449b-3p in the disease are yet to be examined. The present study revealed a distinct downregulation of miR-449b-3p in NSCLC tissue, which was related to the clinicopathological characteristics, and may serve as an independent marker for NSCLC prognosis. NSCLC cell epithelial-mesenchymal transition (EMT), metastasis and migration were distinctly controlled in vitro by miR-449b-3p, that was found to directly target interleukin (IL)-6. Additionally, increased IL-6 level could inhibit miR-449b-3p and suppress the effect of EMT in NSCLC cells by inactivating the Janus kinase 2 (JAK2)/STAT3 signaling pathway. In conclusion, the data from the present study demonstrated that IL-6 is targeted by miR-449b-3p, which affects the JAK2/STAT3 signaling pathway, impacting on the development of NSCLC.

Vascular endothelial growth factor concentration in vitreous humor of patients with severe proliferative diabetic retinopathy after intravitreal injection of conbercept as an adjunctive therapy for vitrectomy.

BACKGROUND: Proliferative diabetic retinopathy (PDR) is a progressive stage of diabetic retinopathy featured by the formation of neovascular and proliferative membrane. Vascular endothelial growth factor (VEGF) acts as a pivot factor in the development of neovascularization. This study was to investigate the changes of intravitreal VEGF concentrations of severe PDR after intravitreal injection of conbercept (IVC) and its potential advantages to the following vitrectomy. METHODS: This was a prospective, interventional, randomized controlled study. Sixty eyes (60 patients) with severe PDR and 20 eyes from 20 patients with rhegmatogenous retinal detachment complicated with proliferative vitreoretinopathy were enrolled in this study. PDR eyes were randomly assigned to three groups by sortation randomization method with 20 eyes in each based on the interval of preoperative IVC (group A: 7 days, group B: 14 days, group C: non-IVC). Another 20 eyes without diabetes were enrolled as the non-diabetic control group (group D), receiving PPV directly. Vitreous specimens of all 80 patients were collected and evaluated afterwards. The intravitreal VEGF concentration of the four groups, and the total surgical time and the intraoperative bleeding rate of the PDR groups were recorded. RESULTS: The mean intravitreal VEGF concentrations of groups A-D were 66.6 ±â€Š43.3, 93.1 ±â€Š52.3, 161.4 ±â€Š106.1 and 1.8 ±â€Š1.2 pg/mL, respectively. It increased significantly in PDR patients (groups A, B and C) (P = 0.002, <0.001, and <0.001, respectively). PDR patients with preoperative IVC (groups A and B) presented significantly lower VEGF concentrations (P < 0.001 and 0.001), intraoperative bleeding rates (P = 0.004) and total surgical time (P < 0.001, P = 0.003) compared with group C. No statistical differences were presented between groups A and B on the three parameters. CONCLUSION: Seven days and 14 days of preoperative IVC are equally efficient and safe for the vitrectomy of severe PDR patients through decreasing vitreous VEGF concentrations, intraoperative bleeding rate and total surgical times.

Circulating factors associated with sarcopenia during ageing and after intensive lifestyle intervention.

BACKGROUND: Ageing, chronic diseases, prolonged inactivity, and inadequate nutrition pose a severe threat to skeletal muscle health and function. To date, experimental evidence suggests that ageing-related subclinical inflammation could be an important causative factor in sarcopenia. Although inflammatory signalling has been implicated in the pathogenesis of experimental animal models of sarcopenia, few studies have surveyed the clinical association between circulating factors and muscle mass in patients before and after lifestyle interventions. In this study, we evaluated whether proinflammatory cytokines are associated with the onset of sarcopenia, which circulating factors are associated with the severity of sarcopenia, and how these factors change after lifestyle interventions in sarcopenic elderly persons. METHODS: A total of 56 elderly subjects (age ≥ 60 years) with sarcopenia and 56 elderly non-sarcopenic subjects, who met entry criteria and had given informed consent, were selected from the Peking Union Medical College Hospital multicentre prospective longitudinal sarcopenia study for testing relevant circulating factors. Thirty-two elderly subjects from the sarcopenic cohort completed a 12 week intensive lifestyle intervention programme with whey supplements (30 g/day) and a personalized resistance training regimen. The levels of proinflammatory cytokines and metabolic hormones, pre-intensive and post-intensive lifestyle interventions, were measured. RESULTS: The sarcopenic group was significantly older (72.05 ± 6.54 years; P < 0.001), more likely to be inactive and female (57.1% of all sarcopenic patients), and had a higher prevalence of type 2 diabetes (16% higher risk). Compared with non-sarcopenic subjects, serum interleukin (IL)-6, IL-18, tumour necrosis factor-α (TNF-α), TNF-like weak inducer of apoptosis (TWEAK), and leptin were significantly higher, while insulin growth factor 1, insulin, and adiponectin were significantly lower in sarcopenic patients (all P < 0.05). Logistic regression analyses revealed that high levels of TNF-α (>11.15 pg/mL) and TWEAK (>1276.48 pg/mL) were associated with a 7.6-fold and 14.3-fold increased risk of sarcopenia, respectively. After adjustment for confounding variables, high levels of TWEAK were still associated with a 13.4-fold increased risk of sarcopenia. Intensive lifestyle interventions led to significant improvements in sarcopenic patients' muscle mass and serum profiles of TWEAK, TNF-α, IL-18, insulin, and adiponectin (all P < 0.05). CONCLUSIONS: High levels of the inflammatory cytokines TWEAK and TNF-α are associated with an increased risk of sarcopenia, while the metabolic hormones insulin growth factor 1, insulin, and adiponectin are associated with a decreased risk of sarcopenia in our Chinese patient cohort. Intensive lifestyle interventions could significantly improve muscle mass, reduce inflammation, and restore metabolic hormone levels in sarcopenic patients. This trial was registered at clinicaltrials.gov as NCT02873676.

Expression of microRNAs-106b in nonsmall cell lung cancer.

AIM: To explore the expression of microRNA-106b (miRNA-106b) in nonsmall cell lung cancer (NSCLC). SETTINGS AND DESIGN: miRNAs are short regulatory RNAs that negatively modulate gene expression at the posttranscriptional level, and are deeply involved in the pathogenesis of several types of cancer. miRNA-106b has been shown to play an oncogenic role in tumor progression. The expression of miRNA-106b is detected in this study. SUBJECTS AND METHODS: Quantitative reverse transcription polymerase chain reaction and Northern blotting were used to detect the expression level of miRNA-106b in 200 NSCLC samples. STATISTICAL ANALYSIS USED: All statistical analyses were performed using SPSS 16.0 software. Results were statistically evaluated using the Kruskal-Wallis test and Mann-Whitney U-test. Survival curves were estimated by the Kaplan-Meier method and P < 0.05 was considered to be statistically significant. RESULTS: miRNA-106b expression is increased in NSCLC tissues. Statistical analysis showed that overexpression of miRNA-106b was strongly associated with lymph node metastasis, stage of tumor node metastasis classification, and poor prognosis. Moreover, there was a significant difference in the miRNA-106b expression levels between smoking and nonsmoking patients. Multivariate Cox regression analysis showed that miRNA-106b was an independent prognostic factor for NSCLC patients. CONCLUSIONS: These data suggest that aberrantly expressed miRNA-106b may contribute to the development of NSCLC.

Antitumor and antiangiogenic activities of anti-vascular endothelial growth factor hairpin ribozyme in human hepatocellular carcinoma cell cultures and xenografts.

AIM: To study the effectiveness and mechanisms of anti- human vascular endothelial growth factor (hVEGF) hairpin ribozyme on angiogenesis, oncogenicity and tumor growth in a hepatocarcinoma cell line and a xenografted model. METHODS: The artificial anti-hVEGF hairpin ribozyme was transfected into hepatocarcinoma cell line SMMC-7,721 and, subsequently, polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR) were performed to confirm the ribozyme gene integration and transcription. To determine the effects of ribozyme ,VEGF expression was detected by semiquantitative RT-PCR and enzyme liked immunosorbent assay (ELISA). MTT assay was carried out to measure the cell proliferation. Furthermore,the transfected and control cells were inoculated into nude mice respectively, the growth of cells in nude mice and angiogenesis were observed. RESULTS: VEGF expression was down-regulated sharply by ribozyme in transfected SMMC-7,721 cells and xenografted tumor. Compared to the control group, the transfected cells grew slower in cell cultures and xenografts, and the xenograft formation was delayed as well. In addition, the microvessel density of the xenografted tumor was obviously declined in the transfected group. As demonstrated by microscopy,reduction of VEGF production induced by ribozyme resulted in a significantly higher cell differentiation and less proliferation vigor in xenografted tumor. CONCLUSION: Anti-hVEGF hairpin ribozyme can effectively inhibit VEGF expression and growth of hepatocarcinoma in vitro and in vivo. VEGF is functionally related to cell proliferation, differentiation and tumori-genesis in hepatocarcinoma.

[Regulation of tyrosylprotein sulfotransferases activity by sulfotyrosine].

OBJECTIVE: To investigate the role of sulfated tyrosine in regulating the activity of tyrosylprotein sulfotransferases (TPST) 1 and TPST2. METHODS: Constructs of TPST 1 and TPST2 were amplified by polymerase chain reaction (PCR), then fused into immunoglobulin G1 Fc region. All the variants in which sulfated tyrosines were mutated to phenylalanine were made by the PCR-based Quick Change method and confirmed by sequencing the entire reading frame. Small hairpin RNA (shRNA) constructs-targeting nucleotides 259-275 of TPST1 and nucleotides 73-94 of TPST2 were generated and subcloned into pBluescript. Human embryonic kidney (HEK) 293T cells were transfected with these plasmids. One day later, cells were split: one part was labeled with 35S-cysteine and methionine or 35S-Na2SO3 overnight, the second part was used for 125I labeled binding experiment, and the third part was retained for binding and flow cytometry. RESULTS: Tyrosines at position 326 of TPST1 and position 325 of TPST2 were sulfated posttranslationally. Tyrosine sulfation of TPSTs was effectively inhibited by sulfation inhibitors, including specific shRNAs and non-specific NaCIO3. shRNAs reduced the sulfation of C3a receptor and C5a receptor, and partially blocked the binding of these two receptors to their respective ligands. CONCLUSIONS: The activities of TPSTs were regulated by tyrosine sulfation. Inhibition of sulfotyrosine decreases the binding ability of C3a receptor and C5a receptor to their respective ligands.

Overexpression of SKP2 Inhibits the Radiation-Induced Bystander Effects of Esophageal Carcinoma.

BACKGROUND: To investigate the effects of S-phase kinase protein 2 (SKP2) expression on the radiation induced bystander effect (RIBE) in esophageal cancer (EC) cells. MATERIALS AND METHODS: Western blot was used to detect the levels of SKP2, Rad51, and Ku70 in EC cells. Positive transfection, RNAi, micronucleus (MN), and γ-H2AX focus formation assay were used to investigate the effects of SKP2 on RIBE induced by irradiated cells. RESULTS: We found a significant negative correlation between SKP2 expression and MN frequency (p < 0.05) induced by RIBE. The results were further confirmed by positive transfection, RNAi, and rescue experiments.γ-H2AX focus formation assay results indicated that overexpression of SKP2 in the irradiated cells inhibited the DNA damage of RIBE cells. However, when SKP2 expression decreased in irradiated cells, the DNA damage of RIBE cells increased. Increased or decreased expression levels of SKP2 had effects on Rad51 expression under the conditions of RIBE. CONCLUSIONS: These results showed, for the first time, that SKP2 expression can inhibit RIBE of EC cells. The mechanism may function, at least partly, through the regulation of Rad51 in the ability to repair DNA damage.

CXC chemokine receptor 3 modulates bleomycin-induced pulmonary injury via involving inflammatory process.

OBJECTIVE: To investigate the role of CXC chemokine receptor 3 (CXCR3) in bleomycin-induced lung injury by using CXCR3 gene deficient mice. METHODS: Sex-, age-, and weight-matched C57BL/6 CXCR3 gene knockout mice and C57BL/6 wide type mice were challenged by injection of bleomycin via trachea. Lung tissue was stained with HE method. Airway resistance was measured. Bronchoalveolar lavage (BAL) was performed using phosphate buffered saline twice, cell number and differentials were counted by Diff-Quick staining. Interleukin (IL)4, IL-5, IL-12p40, and interfon-y in BAL fluid and lung homogenate were measured by enzyme-linked immunosorbent assay. Unpaired t test was explored to compare the difference between two groups. RESULTS: On day 7 after bleomycin injection via trachea, CXCR3 knockout mice were protected from bleomycin-induced lung injury as evidenced by fewer accumulation of inflammatory cells in the airway and lung interstitium compared with their wild type littermates (P < 0.05). Airway resistance was also lower in CXCR3 knockout mice compared with wild type mice (P < 0.01). Significantly lower level of inflammatory cytokines release, including the altered production of IL-4 and IL-5 both in BAL fluid and lung tissue was seen in CXCR3 knockout mice than in wild type mice (both P<0.05). CONCLUSION: CXCR3 signaling promotes inflammatory cells recruiting and initiates inflammatory cytokines cascade following endotracheal bleomycin administration, indicating that CXCR3 might be a therapeutic target for pulmonary injury.

Low dose pirfenidone suppresses transforming growth factor beta-1 and tissue inhibitor of metalloproteinase-1, and protects rats from lung fibrosis induced by bleomycina.

OBJECTIVE: To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 (TGF-beta 1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and matrix metalloproteinase-13 (MMP-13) in lung tissue. METHODS: Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone (25-800 mg x kg(-1) x d(-1)), dexamethasone (3 mg/kg), or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg x kg(-1) x d(-1) at 7 days or 14 days after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in lung tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxyproline. Expression of proteins of TGF-beta 1, TIMP-1, and MMP-13 were detected by Western blotting. RESULTS: At doses of 25, 50, and 100 mg x kg(-1) x d(-1), pirfenidone had significant anti-fibrotic effects for bleomycin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg x kg(-1) x d(-1) (HE: P < 0.01, P < 0.01, and P = 0.064; sirius red: P < 0.05, P < 0.01, and P < 0.05; hydroxyproline: P = 0.595, P < 0.01, and P = 0.976). Pirfenidone at a dosage of 50 mg x kg(-1) x d(-1) inhibited protein expression of TGF-beta1 and TIMP-1 in lung tissue in the early phase (0.79 and 0.75 times of control group), but had no effect on expression of MMP-13. CONCLUSION: Low dose pirfenidone, especially at dosage of 50 mg x kg(-1) x d(-1), has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-beta 1 and TIMP-1 in lung tissue.

[The application of individualized ventilation strategies in acute respiratory distress syndrome].

OBJECTIVE: To explore better ventilation strategies above lower-tidal-volume (LTV) strategy to protect lung function and improve outcome in acute respiratory distress syndrome (ARDS). METHODS: Thirty ARDS patients were enrolled in Department of Critical Care Medicine of Peking Union Medical College Hospital from July, 2004 to June, 2005. They were randomly allocated into two groups, LTV group and individual ventilation (IV) group. Patients received 6 ml/kg tidal volume (V(T)) and high positive end-expiratory pressure (PEEP) in LTV group. In IV group, static pressure-volume (P-V) curve was measured daily, and PEEP and V(T) were set based on P-V variation, and the open-lung potential was evaluated before recruitment maneuvers. The clinical effect, the degree of lung injury and other outcome indicators in two groups were assessed. RESULTS: The mortality rate in 28 days of IV group (35.7%) was lower than that of LVT group (57.2%, chi(2) = 1.265, P > 0.05). The serum surfactant-associated protein D (SP-D) expression in the third and the seventh day of IV group [154 (91 - 217), 149 (91 - 206) mg/L] were higher than those before enrollment [140 (80 - 200) mg/L]; and the IL-8 expression in the third and the seventh day of IV group [179 (122 - 236), 210 (100 - 321) ng/L] were higher than those before enrollment [210 (132 - 289) ng/L]; but all showed no significant difference [chi(2) = 1.265, Z = 1.079, 1.741, -0.879, 0.471, respectively, all P > 0.05]. The free-ICU days in 28 days and free-organ-dysfunction days of IV group [11 (5 - 16) d, 13 (6 - 18) d] were significantly higher than that of LTV group [3 (0 - 8) d, 3 (0 - 7) d, Z = -2.277, -2.372 respectively, all P < 0.05]. The PEEP, V(T), partial pressure of carbon dioxide in arterial blood (PaCO2), the plateau pressure (Pplat) of initial 3 days after enrollment in IV group [(11 +/- 2) cm H2O (1 cm H2O = 0.098 kPa), (511 +/- 66) ml, (37 +/- 5) mm Hg (1 mm Hg = 0.133 kPa), (21 +/- 5) cm H2O] were significant different with those of LTV group [(16 +/- 3) cm H2O, (407 +/- 58) ml, (47 +/- 8) mm Hg, (26 +/- 4) cm H2O, t = -8.019, 6.501, -4.311, -4.823, all P < 0.01]. CONCLUSIONS: Compared with LTV and high PEEP therapy, IV strategies are feasible for decreasing PEEP and Pplat, increasing tidal compliance and V(T), and avoiding CO2 retention. It also increased free-ICU days and free-organ-dysfunction days.

[Expression of mir-106b in esophageal squamous cell carcinoma].

OBJECTIVE: To investigate the expression of mir-106b in esophageal squamous cell carcinoma (ESCC) tissue and analyze its correlation with the clinicopathologic features of ESCC. METHODS: A total of 200 fresh surgical specimens of ESCC and adjacent tissues collected between 2001 and 2007 were examined for expressions of mir-106b using real-time PCR (RT-PCR). Northern blot analysis for mir-106b was performed in 4 pairs of samples to confirm the RT-PCR results. The relationship between mir-106b expression and clinicopathological features and prognosis of the patients were analyzed. RESULTS: Mir-106b was expressed at significantly higher levels in ESCC tissues than in the paired adjacent tissues. Overexpression of mir-106b was associated with lymph node metastasis, stage of TNM classification and smoking (P<0.05). The survival rate of patients with low mir-106b expression was higher than that of patients with high mir-106b expression (60 vs 37 months, P=0.024). Cox regression analysis indicated that the expression of mir-106b, lymph node metastasis and smoking were independent prognostic factors for ESCC (P<0.05). CONCLUSION: Mir-106b is overexpressed in ESCC tumors, and its overexpression is strongly associated with lymph node metastasis and a poor prognosis. Mir-106b expression is an independent prognostic factor for ESCC and can serve as a biomarker for diagnosis and prognostic evaluation of ESCC.

Two luminescent Zn(II) metal-organic frameworks for exceptionally selective detection of picric acid explosives.

Two luminescent Zn(II) metal-organic frameworks were prepared from a π-conjugated thiophene-containing carboxylic acid ligand. These two MOFs show strong luminescene and their luminescence could be quenched by a series of nitroaromatic explosives. Importantly, they exhibit very highly sensitive and selective detection of picric acid compared to other nitroaromatic explosives.