A randomized, controlled, multicenter clinical trial to evaluate the efficacy and safety of oral sitafloxacin versus moxifloxacin in adult patients with community-acquired pneumonia.

OBJECTIVES: To evaluate the efficacy and safety of oral sitafloxacin versus oral moxifloxacin in the treatment of Chinese adults with community-acquired pneumonia (CAP). PATIENTS AND METHODS: This is a multicenter, randomized, open-label, positive-controlled clinical trial (chinadrugtrials.org.cn identifier: CTR20130046). CAP patients received sitafloxacin tablets 100 mg once daily (qd) or 100 mg twice daily (bid) to compare with moxifloxacin tablets 400 mg qd, for 7-10 days. The primary outcome was non-inferiority of sitafloxacin to moxifloxacin in clinical cure rate at test of cure (TOC) visit in per-protocol set (PPS). RESULTS: A total of 343 patients were randomized (sitafloxacin 100 mg qd, n = 117; sitafloxacin 100 mg bid, n = 116; moxifloxacin, n = 110), 291 patients were included in the PPS (sitafloxacin 100 mg qd, n = 96; sitafloxacin 100 mg bid, n = 94; moxifloxacin, n = 101). The clinical cure rate was 94.8% in the sitafloxacin 100 mg qd group, 96.8% in the sitafloxacin 100 mg bid group and 95.0% in the moxifloxacin group. At the TOC visit, the microbiological success rate was 97.0% (32/33) in the sitafloxacin 100 mg qd group, 97.1% (34/35) in the sitafloxacin 100 mg bid group and 94.9% (37/39) in the moxifloxacin group in the microbiological evaluable set (MES). The incidence of study-drug-related adverse events (AEs) was 23.3% (27/116) in the sitafloxacin 100 mg qd group, 29.8% (34/114) in the sitafloxacin 100 mg bid group and 28.2% (31/110) in the moxifloxacin group (p > .05). The common AEs related to study drug were dizziness, nausea, diarrhea, increased platelet count and alanine transaminase (ALT) elevation. All the AEs resolved completely after discontinuation of study drug. CONCLUSION: Sitafloxacin 100 mg qd or 100 mg bid for 7-10 days is not inferior to moxifloxacin 400 mg qd for 7-10 days in clinical efficacy for adult CAP patients. Sitafloxacin provides a safety profile comparable to moxifloxacin.

Baicalin inhibits PDGF-induced proliferation and migration of airway smooth muscle cells.

Airway smooth muscle (ASM) cell proliferation and migration play important roles in airway remodeling in asthma. In vitro platelet-derived growth factor (PDGF) induced ASM cell proliferation and migration. Baicalin is one of flavonoid extracts from Scutellaria baicalensis, which has an anti-asthma effect. However, little is known about its role in PDGF-induced proliferation and migration in rat ASM (RASM) cells. In this study, we aimed to investigate the effects of baicalin on PDGF-induced RASM cell proliferation and migration. We also identified the signaling pathway by which baicalin influences RASM cell proliferation and migration. In the current study, we demonstrated that baicalin suppressed PDGF-induced RASM cell proliferation, arrested PDGF-induced cell-cycle progression. It also suppressed PDGF-induced RASM cell migration. Furthermore, baicalin suppressed PDGF-induced expression of phosphorylated p38, ERK1/2 and JNK in RASM cells. In summary, our study is the first to show that baicalin pretreatment can significantly inhibit PDGF-induced RASM cell proliferation and migration by suppressing the MAPK signaling pathway, and baicalin may be a useful chemotherapeutic agent for asthma.

Impact of TREM-2 gene silencing on inflammatory response of endotoxin-induced acute lung injury in mice.

Acute lung injury (ALI) is one of the critical clinical respiratory diseases, of which infection is the main cause and the first risk factor. This study investigated the impact of triggering receptor of myeloid cells expression (TREM)-2 gene silencing on inflammatory response of endotoxin-induced ALI in mice. Lentivirus-mediated TREM-2-shRNA was transfected into healthy male C57BL/6 mice, and the lipopolysaccharide-induced ALI model was established. The immunohistochemistry, immunofluorescence, fluorescence quantitative PCR, western blot, and ELISA were applied to detect the pathological changes of lung tissue and expressions of TREM-2, tumor necrosis factor-α (TNF-α), and interleukin 10 (IL-10) in bronchoalveolar lavage fluid. The lentivirus group, saline control group, ALI model group, blank control group, and negative control group were set up at the same time. Results found that, in lentivirus group, the pathological change of lung tissue was significantly lighter than ALI model group (P < 0.05), and the expression of TREM-2 was significantly reduced compared with all control groups (P < 0.05). The levels of TNF-α and IL-10 were significantly increased than all control groups (P < 0.05), while above indexes in negative control group and blank control group showed no significant difference with ALI group (P > 0.05). This study indicates that TREM-2 has a protective effect on inflammatory response of endotoxin-induced ALI in mice, which has provided new potential targets for prevention and treatment of ALI.

Matrix metalloproteinases contribute to kidney fibrosis in chronic kidney diseases.

Matrix metalloproteinases (MMPs) are members of the neutral proteinase family. They were previously thought to be anti-fibrotic because of their ability to degrade and remodel of extracellular matrix. However, recent studies have shown that MMPs are implicated in initiation and progression of kidney fibrosis through tubular cell epithelial-mesenchymal transition (EMT) as well as activation of resident fibroblasts, endothelial-mesenchymal transition (EndoMT) and pericyte-myofibroblast transdifferentiation. Interstitial macrophage infiltration has also been shown to correlate with the severity of kidney fibrosis in various chronic kidney diseases. MMPs secreted by macrophages, especially MMP-9, has been shown by us to be profibrotic by induction of tubular cells EMT. EMT is mainly induced by transforming growth factor-ß (TGF-ß). However, MMP-9 was found by us and others to be up-regulated by TGF-ß1 in kidney tubular epithelial cells and secreted by activated macrophages, resulting in EMT and ultimately kidney fibrosis. Therefore, MMP-9 may serve as a potential therapeutic target to prevent kidney fibrosis in chronic kidney disease. This review, by a particular focus on EMT, seeks to provide a comprehensive understanding of MMPs, especially MMP-9, in kidney fibrosis.

Silencing of triggering receptor expressed on myeloid cells-2 enhances the inflammatory responses of alveolar macrophages to lipopolysaccharide.

Triggering receptor expressed on myeloid cells-2 (TREM-2) has been shown to attenuate inflammatory responses in various cell lines including bone marrow-derived macrophages, hepatic macrophages, osteoclasts and dendritic cells. However, its effects on alveolar macrophages remain unknown. Lentivirus-mediated RNA interference (RNAi) is a post-transcriptional gene silencing method, which is capable of degrading target genes specifically and efficiently. In this study, we silenced TREM-2 in murine alveolar macrophages by using lentivirus-mediated short hairpin RNA (shRNA) and evaluated the effects of TREM-2 silencing on expression of toll-like receptor-4 (TLR-4), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in response to lipopolysaccharide (LPS). Alveolar macrophages were transfected with shRNA targeting TREM-2 by use of lentivirus vector, non-sense shRNA as a negative control or empty lentivirus vector as a blank control. Silencing of TREM-2 was assessed by real­time fluorescence quantitative PCR and flow cytometry. Following LPS stimulation, the levels of TLR-4, TNF-α and IL-10 expressed in alveolar macrophages were measured by real-time PCR, flow cytometry or ELISA. TREM-2 expression on alveolar macrophages was downregulated significantly by lentivirus-mediated shRNA treatment at the transcriptional and translational levels. However, alveolar macrophages that received non-sense shRNA or empty lentivirus vectors showed no effects on TREM-2 expression. Silencing of TREM-2 enhanced expression of TLR-4, as well as TNF-α and IL-10, by alveolar macrophages following LPS stimulation. These results indicate a significant effect of TREM-2 on attenuating the LPS-induced inflammatory response of murine alveolar macrophages, which may be dependent on TLR-4.

Effects of storage solutions on the viability of human umbilical cord mesenchymal stem cells for transplantation.

Human umbilical cord-derived mesenchymal stem cell (UC-MSC) transplantation has shown promise for the treatment of various diseases. For clinical applications, UC-MSCs have been stored in 0.9% saline, 5% dextrose, dextrose and sodium chloride injection, Plasma-Lyte A, 1% human serum albumin (1% HSA), or 5% HSA before administration, but the effect of storage conditions on the viability and biological function of the cells remains unknown. Freshly harvested UC-MSCs were resuspended and incubated in these solutions for 2, 4, or 6 h at 4°C or room temperature (24°C). Cell viability, apoptotic/necrotic fraction, poststorage growth potential, immunophenotype, immunosuppressive capacity, and differentiation capacity were analyzed. When stored in parenteral solutions, UC-MSCs showed progressive deterioration in survival viability and adhesion ability. After 6-h storage, the best viability and attachment rate of UC-MSCs decreased to 83.0 ± 1.6% and 71.8 ± 3.2%, respectively. Our results suggested that UC-MSCs in these conditions lose their viability in a short time. However, it seems that the other biological functions of the surviving UC-MSCs were little affected. Since UC-MSCs suspended in these mediums lose their survival viability in a short time to levels significantly below the permissible limits (70%) by FDA, precautions need to be taken on using these solutions as suspension medium and further studies on the optimal methods for preservation are urgent.

[Changes of ophthalmic blood flow in obstructive sleep apnea-hypopnea syndrome].

OBJECTIVE: To observe the ophthalmic artery (OA), central retinal artery (CRA) and posterior ciliary artery (PCA) blood flow and the changes of eye vascular auto-regulation in patients with obstructive sleep apnea-hypopnea syndrome (OSAS). METHODS: It was a case-control study. Fifteen health overweight male adult (as normal control) and 42 patients with OSAS were randomly selected from examination center and sleep detection center, respectively. The OSAS patients were divided into mild (14 patients) and moderate and severe (28 patients) groups based on the apnea-hypopnea index (AHI). All subjects filled the sleep questionnaire and carried out polysomnogram monitoring all night for at least 7 hours. Fasting peripheral venous blood was collected at 7 AM on next day. The end-tidal CO(2) (ETCO(2)), intraocular pressure and color doppler sonography were examined next day to record the data in the inspection process before and after Mueller maneuver. Doppler ultrasound measurement of ocular blood flow diameter and blood flow velocity values were described in the median (max, min) and compared with Kruskal-Wallis test. And then two groups were compared with Bonferroni t test. Ocular blood flow velocity of patients with OSAS and PSG monitoring indicators were analyzed using partial correlation analysis. RESULTS: OA inner diameter in moderate and severe OSAS group [0.08 (0.15, 0.06) cm] was lower than that in healthy control [0.15 (0.26, 0.11) cm] and the difference was statistically significant (P = 0.000). PCA inner diameter in moderate and severe OSAS group [0.10 (0.13, 0.07) cm] were higher than that in healthy controls [0.05 (0.09, 0.04) cm]. CRA peak systolic velocity (PSV) in moderate and severe OSAS group [16.50 (19.40, 13.10) cm/s] was greater than that in healthy controls [11.30 (16.70, 8.20) cm/s]. The differences between these two groups were statistically significant (PCA inner diameter: P = 0.000, CRA-PSV: P = 0.001). The difference of CRA end diastolic velocity (EDV) between the moderate and severe group [8.90 (9.90, 5.10) cm/s], mild group [7.00 (8.30, 4.50) cm/s] and healthy control group [5.50 (7.40, 3.40) cm/s] was statistically significant (χ(2) = 14.45, P < 0.05). PCA-PSV [32.50 (43.10, 19.10) cm/s] and PCA-EDV [12.80 (15.20, 5.70) cm/s] in the moderate and severe group were higher than those in healthy control group [22.60 (32.20, 12.40) cm/s] and [7.20 (11.20, 3.90) cm/s], as well as those in the mild group [24.00 (30.70, 13.30) cm/s] and [8.00 (9.90, 3.90) cm/s]. These differences were statistically significant (PCA-PSV: P = 0.000, 0.002; PCA-EDV: P = 0.000, 0.001). The diameter of OA and PCA correlated negatively with ETCO(2) (r = -0.41, -0.34; P < 0.05); CRA-PSV was correlated with SaO2 min (r = -0.37, P < 0.05). CRA-EDV was correlated with ETCO(2) and SaO2 mean (r = 0.57, -0.39; P < 0.05). PCA-PSV was correlated with SaO2 min and MAI (r = -0.34, 0.56; P < 0.05). PCA-EDV was correlated SaO2 min and MAI (r = -0.29, 0.61; P < 0.05). CONCLUSIONS: The diameter and blood flow of OA, PCA and CRA change in OSAS patients. Compared with non-OSAS patients, the autoregulation function of PCA and CRA is weakened in OSAS patients.

E-cadherin/ß-catenin complex and the epithelial barrier.

E-Cadherin/ß-catenin complex plays an important role in maintaining epithelial integrity and disrupting this complex affect not only the adhesive repertoire of a cell, but also the Wnt-signaling pathway. Aberrant expression of the complex is associated with a wide variety of human malignancies and disorders of fibrosis resulting from epithelial-mesenchymal transition. These associations provide insights into the complexity that is likely responsible for the fibrosis/tumor suppressive action of E-cadherin/ß-catenin.

Inhibition of allergic airway inflammation by antisense-induced blockade of STAT6 expression.

BACKGROUND: The signal transducer and activator of transcription 6 (STAT6) expression in lung epithelial cells plays a pivotal role in asthma pathogenesis. Activation of STAT6 expression results in T helper cell type 2 (Th2) cell differentiation leading to Th2-mediated IgE production, development of allergic airway inflammation and hyperreactivity. Therefore, antagonizing the expression and/or the function of STAT6 could be used as a mode of therapy for allergic airway inflammation. METHODS: In this study, we synthesized a 20-mer phosphorothioate antisense oligonucleotide (ASODN) overlapping the translation starting site of STAT6 and constructed STAT6 antisense RNA (pANTI-STAT6), then transfected them into murine spleen lymphocytes and analyzed the effects of antagonizing STAT6 function in vitro and in a murine model of asthma. RESULTS: In vitro, we showed suppression of STAT6 expression and interleukin (IL)-4 production of lymphocytes by STAT6 ASODN. This effect was more prominent when cells were cultured with pANTI-STAT6. In a murine model of asthma associated with allergic pulmonary inflammation in ovalbumin (OVA)-sensitized mice, local intranasal administration of fluorescein isothiocyanate (FITC)-labeled STAT6 ASODN to DNA uptake in lung cells was accompanied by a reduction of intracellular STAT6 expression. Such intrapulmonary blockade of STAT6 expression abrogated signs of lung inflammation, infiltration of eosinophils and Th2 cytokine production. CONCLUSION: These data suggest a critical role of STAT6 in the pathogenesis of asthma and the use of local delivery of STAT6 ASODN as a novel approach for the treatment of allergic airway inflammation such as in asthma.

[The association between polymorphism of beta(1)-adrenoceptor genes and cardiovascular complication in obstructive sleep apnea-hypopnea syndrome patients in Han population].

OBJECTIVE: To investigate the association between polymorphism of beta(1)-adrenoceptor genes and cardiovascular complication in obstructive sleep apnea-hypopnea syndrome (OSAHS) patients. METHOD: The Gly389Arg polymorphism of beta(1)-adrenoceptor genes was identified by polymerase chain reaction-restricted fragment length polymorphism assay in 192 OSAHS patients of "Han" population of China. The effect of polymorphisms in the OSAHS group on parameters of polysomnography and cardiovascular complication was analyzed. Analysis of variance, t test and chi-square test were used for statistics. RESULT: The distributions with CC genotype were significantly higher than those with CG/GG genotype. The frequencies with C allele were significantly higher than those with G allele. The OSAHS patients with CC genotype had higher percentage of time of oxygen saturation lower than 90% (T90) and longer apnea time (Tmax) than those with CG/GG genotype. The lowest saturation of blood oxygen (minSaO2) in patients with CC genotype was significantly lower than that in patients with CG/GG genotype. Cardiovascular diseases (P < 0.05) except sinus bradycardia were more common in patients with the CC genotype(P < 0.05). CONCLUSION: The Gly389Arg CC genotype of beta(1)-adrenoceptor genes may be involved in cardiovascular complication in OSAHS, and the C allele may be an important candidate gene.

[Restoration of pathological changes of emphysema by angiogenesis factors: experiment of rats].

OBJECTIVE: To investigate whether intratracheal administration of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) restore the pulmonary function and pathology in emphysema, and research the mechanism of they restored pulmonary emphysema, and the pathogenesis of pulmonary emphysema. METHODS: Twenty-four Wistar rats were randomized into the 4 equal groups: bFGF group [receiving a single intratracheal instillation of porcine pancreatic elastase (PPE) 250 U/kg, and 4 weeks later receiving intratracheal instillation of bFGF 400 U once a week for 3 weeks), VEGF group (receiving a single intratracheal instillation of PPE 250 U/kg, and 4 weeks later receiving intratracheal instillation of VEGF 2 microg once a week for 3 weeks), control group [receiving a single intratracheal instillation of PPE 250 U/kg, and 4 weeks later receiving intratracheal instillation of normal saline (NS) once a week for 3 weeks], and normal group (receiving intratracheal instillation of NS in above-mentioned pattern). Four weeks after treatment, arterial blood sample was collected from the abdominal aorta to undergo blood gas analysis for assessment pulmonary function, and then the rata were killed with their lungs taken out to undergo pathological examination. Immunohistochemistry was performed to detect the CD34, markers of pulmonary capillary endothelial cells. RESULTS: There were no significant differences in the artery blood gas analysis among the four groups (all P > 0.05). The levels of mean alveoli number (MAN) of the bFGF and VEGF groups were (43 +/- 8)/HP and (44 +/- 9)/HP] respectively, both significantly higher than that of the control group [(30 +/- 6)/HP, both P < 0.01]. The levels of mean linear intercept (MLI) of the bFGF and VEGF groups were (196 +/- 38) microm and (194 +/- 38) microm respectively, both significantly lower than that of the control group [(288 +/- 68) microm, both P < 0.01). the mean alveoli area (MAA) level of the bFGF and VEGF groups were (9856 +/- 1864) microm(2) and (9804 +/- 1929) microm(2) respectively, both significantly lower than that of the control group [(14,525 +/- 3408) microm(2), both P < 0.01]. The percentages of CD34(+) cells of the bFGF and VEGF groups were (3.7 +/- 1.3)% and (2.6 +/- 1.2)% respectively, both significantly higher than that of the control group [(0.8 +/- 0.7)%, both P < 0.05). CONCLUSION: bFGF and VEGF can restore the pathological changes of experimental emphysema. The damage of pulmonary capillary may play an important role in the pathogenesis of pulmonary emphysema.

[The effects of bilirubin concentration on laminin and epidermal growth factor expression in lung tissue and type II pneumocytes in smoking rats model].

OBJECTIVE: To explore the effects of bilirubin on expression of laminin (Ln) and epidermal growth factor (EGF) in lung tissue and in type II pneumocytes (ATII) in smoking rats. METHODS: 36 healthy Wistar rats were randomly divided into three groups: a normal group, a smoking-induced emphysema model group (model group), and a bilirubin group (n = 12, for each group). According to XU San-lin method, rats both in the model group and the bilirubin group were exposed to smoke for a total of 6 months. Rats in the bilirubin group were pretreated with indirect bilirubin (20 mg.kg(-1).d(-1)) before exposed to smoke once a day. All animals were sacrificed six months later. ATIIwere isolated, purified and cultured. Ln in ATII culture medium was determined by radioimmunoassay (RIA). We detected Ln in lung tissue with indirect immunofluorescence technique and EGF with immunochemistry. RESULTS: In the model group, Ln content in ATII culture medium and in lung tissue [(5.0 +/- 0.3) mg/ml, 5.67 +/- 0.26] increased significantly compared with those in the normal group [(3.2 +/- 0.5) mg/ml, 2.01 +/- 0.74; P < 0.05, respectively]. In the bilirubin group, the two indexes [(3.9 +/- 0.7) mg/ml, 4.18 +/- 0.63] decreased significantly compared with those in the model group (P < 0.05, respectively), which was higher than the normal group (P < 0.05). Expression of EGF in lung tissue of the model group (0.43 +/- 0.09) increased significantly compared with that of the normal group (0.14 +/- 0.02; P < 0.05). The expression of EGF in the bilirubin group (0.34 +/- 0.03) decreased significantly compared with the model group (P < 0.05), which was higher than that of the normal group. CONCLUSIONS: Bilirubin was shown to be able to promote the reconstruction of extracellular matrix by decreasing the expression of Ln and EGF in lung tissue and in ATII in the development of emphysema.

[Epidemiological studies of sleep apnea-hypopnea syndrome: a questionnaire survey of Taiyuan].

OBJECTIVE: To obtain the epidemiological data of sleep apnea-hypopnea sydrome (SAHS) in Taiyuan. METHODS: A questionnaire survey was performed in 6 028 people living in Taiyuan. The prevalence of SAHS was estimated by a two-stage procedure. In the first stage, stratified cluster disproportional random sampling survey was performed in Taiyuan. 6 028 questionnaires were send to random sample of defined population in the 4 sites selected from 2 districts. The response rate was 85.11%. During the second stage 476 of those highly suspected of having SAHS (ESS >/= 9) underwent all-night polysomnographic (PSG) studies. RESULTS: From the study population, 179 were diagnosed as having SAHS. The overall prevalence was 3.5% (male 4.7% and female 1.9%). CONCLUSION: The prevalence of SAHS was 3.5% among Taiyuan.