Molecular motor tug-of-war regulates elongasome cell wall synthesis dynamics in Bacillus subtilis.

Most rod-shaped bacteria elongate by inserting new cell wall material into the inner surface of the cell sidewall. This is performed by class A penicillin binding proteins (PBPs) and a highly conserved protein complex, the elongasome, which moves processively around the cell circumference and inserts long glycan strands that act as barrel-hoop-like reinforcing structures, thereby giving rise to a rod-shaped cell. However, it remains unclear how elongasome synthesis dynamics and termination events are regulated to determine the length of these critical cell-reinforcing structures. To address this, we developed a method to track individual elongasome complexes around the entire circumference of Bacillus subtilis cells for minutes-long periods using single-molecule fluorescence microscopy. We found that the B. subtilis elongasome is highly processive and that processive synthesis events are frequently terminated by rapid reversal or extended pauses. We found that cellular levels of RodA regulate elongasome processivity, reversal and pausing. Our single-molecule data, together with stochastic simulations, show that elongasome dynamics and processivity are regulated by molecular motor tug-of-war competition between several, likely two, oppositely oriented peptidoglycan synthesis complexes associated with the MreB filament. Altogether these results demonstrate that molecular motor tug-of-war is a key regulator of elongasome dynamics in B. subtilis, which likely also regulates the cell shape via modulation of elongasome processivity.

LINC00346 regulates NLRP1-mediated pyroptosis and autophagy via binding to microRNA-637 in vascular endothelium injury.

Endothelial injury and dysfunction contributes to atherosclerosis. LINC00346 plays a key role in vascular endothelial cell injury, however, the specific mechanism remains unclear. This study intends to further explore the relationship between LINC00346 and vascular endothelial injury. Circulating LINC00346 was significantly elevated in patients with coronary artery disease and had high diagnostic value for coronary artery disease. In cell experiments, we found that LINC00346 expression was significantly increased in the oxidized low-density lipoprotein (ox-LDL) intervention group, and LINC00346 knockdown delayed ox-LDL induced human umbilical vein endothelial cell (HUVEC) endothelial-to-mesenchymal transition. In addition, knockdown of LINC00346 mitigated ox-LDL-induced NOD-like receptor protein 1 (NLRP1)-mediated inflammasome formation and pyroptosis, but had no significant effect on NLRP3. By observing the number of autophagosome and detecting intracellular autophagic flux, we found that LINC00346 knockdown inhibited the ox-LDL-induced increase in intracellular autophagy level. Dual-luciferase reporter assay, RNA immunoprecipitation assay, and RNA-pull down assay were performed to confirm the inter-molecular interaction. LINC00346 acted as microRNA-637 sponge to up-regulate the expression of NLRP1. Up-regulation of microRNA-637 alleviated NLRP1-mediated pyroptosis in HUVEC and reduced intracellular autophagosome and autolysosome formation. Finally, we explored whether pyropotosis and autophagy interact with each other. We found that inhibition of intracellular autophagy could alleviate NLRP1-mediated pyroptosis. In conclusion, LINC00346 inhibited the activation of NLRP1-mediated pyroptosis and autophagy via binding to microRNA-637, therefore mitigating vascular endothelial injury.

Immunosuppressive effects of dimethyl fumarate on dendritic cell maturation and migration: a potent protector for coronary heart disease.

Dendritic cells (DCs), as a bridge between innate and adaptive immunity, play key roles in atherogenesis, particularly in plaque rupture, the underlying pathophysiologic cause of myocardial infarction. Targeting DC functions, including maturation and migration to atherosclerotic plaques, may be a novel therapeutic approach to atherosclerotic disease. Dimethyl fumarate (DMF), an agent consisting of a combination of fumaric acid esters, in current study were found to be able to suppress DC maturation by reducing the expression of costimulatory molecules and MHC class II and by blocking cytokine secretion. In addition, DMF efficiently inhibited the migration of activated DCs in vitro and in vivo by reducing the expression of chemokine receptor 7 (CCR7). Additionally, DMF efficiently inhibited the expression of the costimulatory molecule CD86, as well as the chemokine receptor CCR7 and the C-X-C motif chemokine receptor 4 (CXCR4), in healthy donor-derived purified DCs that had been stimulated by ST-segment elevation myocardial infarction (STEMI) patient serum. This study points to the potent therapeutic value of DMF for protecting against cardiovascular disease by suppressing DC functions.

KLF5/LINC00346/miR­148a­3p axis regulates inflammation and endothelial cell injury in atherosclerosis.

Atherosclerosis (AS) is the main pathological basis of cardiovascular diseases, which are related to high morbidity and mortality rates. The present study aimed to investigate the role of the Krüppel­like factor 5 (KLF5)/LINC00346/miR­148a­3p loop in AS. The expression levels of KLF5 in serum and of KLF5/LINC00346/miR­148a­3p in human umbilical vein endothelial cells (HUVECs) were detected by RT­qPCR analysis. The protein expression levels of KLF5, phosphorylated (p­)endothelial nitric oxide synthase (eNOS) and eNOS in HUVECs were analyzed by western blot analysis. Changes in the levels of TNF­α, IL­1ß, IL­6 and nitric oxide (NO) were determined in the supernatant through the application of available commercial kits. The binding of KLF5 to the promoter region of LINC00346 was verified by chromatin immunoprecipitation (ChIP)­PCR assay. The combinatory interaction between KLF5 and LINC00346, LINC00346 and miR­148a­3p, and miR­148a­3p and KLF5 was confirmed by luciferase reporter assay. The results revealed that KLF5 expression was increased in the serum of patients with AS and also in oxidized low­density lipoprotein (OX­LDL)­stimulated HUVECs. The transcription factor KLF5 promoted the transcription of LINC00346. KLF5 interference or LINC00346 interference inhibited the expression of inflammatory factors and functional injury in OX­LDL­stimulated HUVECs. LINC00346 functioned as a sponge of miR­148a­3p. miR­148a­3p overexpression inhibited the expression of inflammatory factors and functional injury in OX­LDL­stimulated HUVECs and miR­148a­3p targeted KLF5 expression. On the whole, the present study demonstrates that KLF5 interference induces the downregulation of LINC00346 and also inhibits inflammation and functional injury in OX OX­LDL­stimulated HUVECs by upregulating miR­148a­3p expression.

Effects of Chinese Herbal Medicine on Acute Exacerbations of COPD: A Randomized, Placebo-Controlled Study.

Purpose: Acute exacerbation of chronic obstructive pulmonary disease (AECOPD) is an essential occurrence in COPD management and is the leading cause of morbidity and mortality. Chinese herbal medicine is widely used in the treatment of AECOPD, but high quality randomized controlled trials are limited. This study aimed to evaluate the efficacy and safety of Chinese herbal medicine as adjuvant therapy for patients with AECOPD. Methods: This was a randomized, double-blind, placebo-controlled study of 378 participants from eight centers in China. Participants were randomly assigned to receive 10 g of Chinese herbal medicine (according to the type of Traditional Chinese medicine syndrome: Sanhanhuayin, Qingrehuatan, or Zaoshihuatan granules) or placebo, two times per day, for 14 days, in addition to conventional medicine. Participants were followed up for 84 days after the treatment. The primary end point was the COPD assessment test (CAT) score. Secondary end points included the Modified British Medical Research Council (mMRC) questionnaire and the COPD patient-reported outcome scale (COPD-PRO). We also assessed treatment failure and treatment success rate, length of hospitalization, number of patients with acute exacerbations, number of patients readmitted due to AECOPD, and number of deaths and intubation. Results: The between-group difference in the change from baseline for CAT on day 14 (end of treatment) was -2.11 (95% confidence interval [CI], -3.198 to -1.050; P<0.001), exceeding the minimal clinically important difference. The mMRC and COPD-PRO scores were lower in the intervention group compared to the control group (between-group difference in the change from baseline, -0.28; 95% CI, -0.48 to -0.08; P=0.007 and -2.51; 95% CI, -4.087 to -0.929; P=0.002, respectively) on day 14. The intervention group had a significantly shorter duration of hospital stay than the control group (mean difference, -1.21days; 95% CI, -2.041 to -0.419; P=0.003), significantly lower of number of exacerbations (risk ratio [RR], 0.60; 95% CI, 0.409 to 0.892; P=0.010), and significantly lower number of readmissions due to AECOPD (RR, 0.41; 95% CI, 0.193 to 0.865; P=0.015). Significant differences in the number of treatment failures or successes, deaths, and intubation were not observed. The difference in safety variables and adverse events between the two groups was not observed. Conclusion: Chinese herbal medicine appears to be safe and beneficial for AECOPD and can be considered a complementary treatment for patients with AECOPD.

The effective evaluation on symptoms and quality of life of chronic obstructive pulmonary disease patients treated by comprehensive therapy based on traditional Chinese medicine patterns.

OBJECTIVE: To evaluate the efficacy of comprehensive interventions based on the three TCM patterns on symptoms and quality of life of COPD patients. DESIGN: An open-label, randomized, controlled trial. SETTING: Four hospitals in China. INTERVENTION: 352 patients were randomly divided into two groups. Patients in the trial group were given conventional Western medicine and Bu-Fei Jian-Pi granules, Bu-Fei Yi-Shen granules and Yi-Qi Zi-Shen granules respectively; patients in the control group were given conventional Western medicine. Data collection was performed at baseline, in the 3rd and 6th month during the treatment period, and the 12th month during the follow-up period. OUTCOMES: Symptoms, including cough, sputum, pant, chest tightness, short of breath, lassitude, cyanosis and symptom total score; quality of life, measured by the WHOQOL-BREF questionnaire and adult COPD quality of life questionnaire (COPD-QOL). RESULTS: Of the 352 patients, 306 fully completed the study. After treatment and follow-up, there were significant differences between two groups in the following: cough, sputum, pant, chest tightness, shortness of breath, lassitude score and symptom total score (P<.05); physical, psychological, social and environment domain (P<.05) of the WHOQOL-BREF; daily living ability, social activity, depression symptoms and anxiety symptoms domain (P<.05) of the COPD-QOL. There were no differences between two groups in cyanosis and adverse events. CONCLUSION: Based on the TCM patterns, Bu-Fei Jian-Pi granules, Bu-Fei Yi-Shen granules and Yi-Qi Zi-Shen granules have beneficial effects on symptoms and quality of life of COPD patients.

Intravenous N-acetylcysteine for prevention of contrast-induced nephropathy: a meta-analysis of randomized, controlled trials.

BACKGROUND: Contrast-induced nephropathy (CIN) is one of the common causes of acute renal insufficiency after contrast procedures. Whether intravenous N-acetylcysteine (NAC) is beneficial for the prevention of contrast-induced nephropathy is uncertain. In this meta-analysis of randomized controlled trials, we aimed to assess the efficacy of intravenous NAC for preventing CIN after administration of intravenous contrast media. STUDY DESIGN: Relevant studies published up to September 2012 that investigated the efficacy of intravenous N-acetylcysteine for preventing CIN were collected from MEDLINE, OVID, EMBASE, Web of Science, Cochrane Central Register of Controlled Trials, and the conference proceedings from major cardiology and nephrology meetings. The primary outcome was CIN. Secondary outcomes included renal failure requiring dialysis, mortality, and length of hospitalization. Data were combined using random-effects models with the performance of standard tests to assess for heterogeneity and publication bias. Meta-regression analyses were also performed. RESULTS: Ten trials involving 1916 patients met our inclusion criteria. Trials varied in patient demographic characteristics, inclusion criteria, dosing regimens, and trial quality. The summary risk ratio for contrast-induced nephropathy was 0.68 (95% CI, 0.46 to 1.02), a nonsignificant trend towards benefit in patients treated with intravenous NAC. There was evidence of significant heterogeneity in NAC effect across studies (Q = 17.42, P = 0.04; I(2) = 48%). Meta-regression revealed no significant relation between the relative risk of CIN and identified differences in participant or study characteristics. CONCLUSION: This meta-analysis showed that research on intravenous N-acetylcysteine and the incidence of CIN is too inconsistent at present to warrant a conclusion on efficacy. A large, well designed trial that incorporates the evaluation of clinically relevant outcomes in participants with different underlying risks of CIN is required to more adequately assess the role for intravenous NAC in CIN prevention.

Effects of comprehensive therapy based on traditional Chinese medicine patterns in stable chronic obstructive pulmonary disease: a four-center, open-label, randomized, controlled study.

BACKGROUND: Traditional Chinese medicine (TCM) has been used to treat chronic obstructive pulmonary disease (COPD) for many years. This study aimed to evaluate the efficacy and safety of the comprehensive therapy based on the three common TCM patterns in stable COPD patients. METHODS: A four-center, open-label randomized controlled method was conducted. A total of 352 patients were divided into the trial group (n = 176, treated with conventional Western medicine and Bu-Fei Jian-Pi granules, Bu-Fei Yi-Shen granules, and Yi-Qi Zi-Shen granules based on the TCM patterns respectively) and the control group (n = 176, treated with conventional Western medicine). The frequency and duration of acute exacerbation, lung function, clinical symptoms, 6-minute walking distance (6MWD), dyspnea scale and quality of life were observed during a 6-month treatment period and at a further 12-month follow-up. RESULTS: A total of 306 patients completed the study fully. The full analysis set (FAS) population was 350 and the per-protocol analysis set (PPS) population was 306. After the 6-month treatment and 12-month follow-up, there were significant differences between the trial and control group in the following: frequency of acute exacerbation (FAS: P = 0.000; PPS: P = 0.000); duration of acute exacerbation (FAS: P = 0.000; PPS: P = 0.001); FEV1 (FAS: P = 0.007; PPS: P = 0.008); symptoms (FAS: P = 0.001; PPS: P = 0.001); 6MWD (FAS: P = 0.045; PPS: P = 0.042); dyspnea scale (FAS: P = 0.002; PPS: P = 0.004); and physical domain (FAS: P = 0.000; PPS: P = 0.000), psychological domain (FAS: P = 0.008; PPS: P = 0.011), social domain (FAS: P = 0.001; PPS: P = 0.000) and environment domain (FAS: P = 0.015; PPS: P = 0.009) of the WHOQOL-BREF questionnaire. There were no differences between the trial and control group in FVC, FEV1% and adverse events. CONCLUSIONS: Based on the TCM patterns, Bu-Fei Jian-Pi granules, Bu-Fei Yi-Shen granules and Yi-Qi Zi-Shen granules have beneficial effects on measured outcomes in stable COPD patients over the 6-month treatment and 12-month follow-up, with no relevant between-group differences in adverse events. TRIAL REGISTRATION: This trial was registered at Chinese Clinical Trial Register Center, ChiCTR-TRC-11001406.

Prognostic factors for community-acquired pneumonia in middle-aged and elderly patients treated with integrated medicine.

OBJECTIVE: To identify prognostic factors in middle-aged and elderly patients with community-acquired pneumonia (CAP) who underwent integrated interventions involving traditional Chinese medicine (TCM) and modern medicine. METHODS: Patients aged > or =45 years and diagnosed with CAP were divided into a middle-aged cohort (45-59 years) and an elderly cohort (> or =60 years), and clinical data comprising 75 predictor variables in seven classes were collected. After replacing missing data, calibrating multicenter differences and classifYing quantitative data, univariate and multivariate analysis were performed. RESULTS: On multivariate analysis, eight independent risk factors--respiration rate, C reactive protein (CRP), cost of hospitalization, anemia, gasping, confusion, moist rales and pneumonia severity index (PSI)--were correlated with the outcome "not cured" in the elderly cohort. Nine factors--neutrophil percentage (Neu%), blood urea nitrogen (BUN), time to clinical stability, appetite, anemia, confusion, being retired or unemployed, Gram-negative bacterial infection and educational level were correlated with not cured in the middle-aged cohort. CONCLUSION: Independent predictive risk factors correlated with adverse outcomes in elderly patients were higher respiration rate, CRP > or = four times the mean or median for the patient's center, cost of hospitalization >11,323 RMB and PSI >11, plus anemia, gasping, confusion and moist rales; those in middle-aged patients were higher Neu%, BUN > or = mean or median, loss of appetite, anemia, confusion, being retired or unemployed and lower educational level. Gram-negative bacterial infection and time to clinical stability >9 days were protective factors.

[Peripheral dendritic cell subsets and changes in plasma monocyte chemoattractant protein 1 in patients with coronary heart disease].

OBJECTIVE: To assess the association between peripheral blood dendritic cells subtype distribution and plasma monocyte chemoattractant protein 1 (MCP-1) concentration in patients with coronary heart disease (CHD). METHODS: Sixty consecutive CHD patients admitted in our department during the period from November, 2010 to December, 2011 were enrolled, including 10 with stable angina pectoris (SAP), 25 with unstable angina pectoris (UAP), and 25 with acute myocardial infarction (AMI), with 28 healthy volunteers as normal controls. All the subjects underwent routine tests and coronary angiography. The percentages of peripheral blood myeloid dendritic cells (mDCs) and plasma cell-like dendritic cells (pDCs) in peripheral blood mononuclear cells were detected by flow cytometry, and plasma MCP-1 levels were detected using enzyme-linked immunosorbent assay. RESULTS: The percentage and absolute quantity of mDCs and pDCs were significantly lower in AMI and UAP groups than in the normal control and SAP groups (P<0.001). In the CHD patients, the plasma MCP-1 level was significantly higher than that in the normal control group (P<0.001) with an inverse correlation with the percentage of peripheral mDCs. CONCLUSION: MCP-1 may promote the migration of mDCs into atherosclerotic plaques and mediate the local immune and inflammatory responses to aggravate plaque instability in CHD patients.

Randomized controlled multicenter clinical trial for integrated treatment of community-acquired pneumonia based on traditional Chinese medicine syndrome differentiation.

OBJECTIVE: To evaluate the efficacy and safety of treatment based on syndrome differentiation of Traditional Chinese Medicine (TCM) for community-acquired pneumonia (CAP). METHODS: A total of 240 CAP patients were randomly divided into the following two groups: the control group was treated by anti-infection plus conventional medicine treatment; and the trial group was treated by TCM plus the above-mentioned treatment given to the controls. The course of treatment was 14 days, and the patients were followed up for 7 days. RESULTS: Of the 240 patients, 235 accomplished the whole process of treatment. The five patients who withdrew from the study were brought into an intent-to-treat analysis. The therapeutic effects of the trial group were superior to those of the control group (P < 0.01). The trial group took less time to become clinically stable, with a higher score in the quality of life (P < 0.01). There were no significant differences in mortality rate (P > 0.05), white blood cell count (P > 0.05), bacterial clearance rate (P > 0.05), and adverse reactions between the two integrated groups. CONCLUSION: Treatment based on TCM syndrome differentiation for CAP has the advantages of resulting in less time to achieve a stable clinical condition, improvement of clinical symptoms and quality of life, and is comparatively safe.

[CCERW: a new rule to predict treatment failure in patients with community-acquired pneumonia in middle aged and elderly people].

OBJECTIVE: To develop and validate a clinical rule to predict treatment failure in middle-aged and elderly patients suffering from community-acquired pneumonia (CAP) in China, and to compare it with other prognostic rules. METHODS: Data of 58 variables affiliated to 6 aspects, including demographics, underlaying diseases, previous status, complications, symptoms, signs and laboratory examination results from the CAP patients aged≥45 years admitted to the respiratory departments in three university affiliated hospitals between December 17, 2006 and December 22, 2008 were enrolled prospectively and then validated in two groups to create a derivation cohort with 75% of the patients for rule development and an internal validation cohort with the other 25% for internal test. An external validation cohort was formed at the same time with patients admitted to the other university affiliated hospital for external test. The single outcome was treatment failure at the time of 14 days after admitted or at discharge from hospital. Univariate analysis, multivariate analysis and receiver operating characteristics (ROC) curve were used for the rule establishment, assessment and comparison among the pneumonia severity index (PSI), CURB65 [confusion, blood urea nitrogen>6.8 mmol/L, respiratory rate (RR)≥30 breaths per minute, systolic blood pressure (SBP)<90 mm Hg (1 mm Hg=0.133 kPa) or diastolic blood pressure (DBP)≤60 mm Hg, age≥65 years] and CRB65 (confusion, RR≥30 breaths per minute, SBP<90 mm Hg or DBP ≤60 mm Hg, age≥65 years). RESULTS: The data of a total of 539 patients in three hospitals were enrolled for analysis. Of those, 400 and 139 patients were randomly allocated into the derivation cohort or internal validation cohort respectively. Meanwhile, 159 patients were enrolled in the external validation cohort. Analyzing the data from 400 patients in the derivation cohort, 33 variables of 6 aspects had significant difference between cure and treatment failure outcome in the univariate analysis. Then, in the multivariate analyses, five independent predictive factors showed significant difference, including confusion (C), creatinine<60 µmol/L, electrolyte disturbances (E), respiratory failure (R), white blood cell (WBC)>7.5×10(9)/L. A clinical prediction rule CCERW based on these variables showed that the treatment failure outcome increased directly with increasing scores: 5.5%-9.1%, 12.8%-20.0% and 31.0%-40.5% for scores of 0-1, 2 and 3-6, respectively. ROC curve analysis yielded an area under the curve (AUC) for CCERW of 0.709 [95% confidence intervals (95%CI) 0.638-0.780], 0.725 (95%CI 0.613-0.838) and 0.686 (95%CI 0.590-0.782) in the derivation, internal and external validation cohorts respectively; and in the same manner, of 0.710 (95%CI 0.659-0.761) for total 698 patients, which was better than PSI, CURB65 and CRB65, at 0.667 (95%CI 0.614-0.719), 0.648 (95%CI 0.592-0.705), and 0.584 (95%CI 0.530-0.638), respectively. CONCLUSION: CCERW can help physicians to distinguish high and low risk leading to treatment failure in middle-aged and elder patients with CAP, and has better predictable capability than PSI, CURB65 and CRB65. We prudent recommend the simple rule can be used in the middle-aged and elder patients with CAP of Han race people in China.