Exosomes Derived from Endothelial Cells Inhibit Neointimal Hyperplasia Induced by Carotid Artery Injury in Rats via ROS-NLRP3 Inflammasome Pathway.

This study attempted to investigate whether exosomes derived from rat endothelial cells (EC-Exo) attenuate intimal hyperplasia after balloon injury using hematoxylin and eosin staining, immunohistochemistry, immunofluorescence staining, Evans blue staining, and Western blotting. The results indicated that EC-Exo inhibited intimal hyperplasia in the carotid artery after balloon injury, promoted re-endothelialization, and reduced vascular inflammation and ROS-NLRP3-mediated cell pyroptosis. Thus, EC-Exo can inhibit neointimal hyperplasia after carotid artery injury in rats presumably by inhibiting the ROS-NLRP3 inflammasome and phenotypic transformation of vascular smooth muscle cells.

Extracorporeal membrane oxygenation (ECMO) in patients with COVID-19: a rapid systematic review of case studies.

OBJECTIVE: COVID-19 can cause severe acute respiratory distress syndrome (ARDS). Extracorporeal membrane oxygenation (ECMO) can support gas exchange in patients failing conventional mechanical ventilation, but its role is still controversial. We performed a rapid systematic review focusing on the use of ECMO in patients with COVID-19. MATERIALS AND METHODS: PubMed/MEDLINE, Google Scholar, Embase, the Cochrane Library, EBSCO and Ovid (updated 30 April 2020) were systematically searched. Case reports/Case series from COVID-19 patients treated with ECMO were included in the study. Three reviewers assessed, selected, and abstracted data from studies. All disparate opinions were resolved through discussion. RESULTS: We included 13 articles for systematic evaluation, including 10 case reports and 3 case series studies, with a total of 72 patients. We search for the following information: First author of articles; Patient's location; age; gender; body mass index (BMI); Comorbidities; Time on ECMO; Mode of ECMO; treatments and clinical outcomes. As of all reporting times, our data show that 38 patients (52.8%) have died definitively, 13 patients (18.0%) were still receiving ECMO treatment, 12 patients (16.7%) were alive, 7 patients (9.7%) were recovery and 2 cases (2.8%) remained hospitalized. CONCLUSIONS: ECMO plays an important role in the stabilization and survival critically ill patients with COVID-19, but the usefulness of ECMO in reducing the mortality of severe ARDS caused by COVID-19 was limited. Therefore, a larger sample size study and a comprehensive analysis of evaluating the medical value of using ECMO on COVID-19 patients are urgently required.

Adjuvant chemotherapy for patients with gastric neuroendocrine carcinomas or mixed adenoneuroendocrine carcinomas.

BACKGROUND: The aim of this study was to evaluate whether adjuvant chemotherapy is associated with improved survival in patients with resectable gastric neuroendocrine carcinomas (G-NECs) or mixed adenoneuroendocrine carcinomas (G-MANECs). METHODS: The study included patients with G-NECs or G-MANECs who underwent surgery in one of 21 centres in China between 2004 and 2016. Propensity score matching analysis was used to reduce selection bias, and overall survival (OS) in different treatment groups was estimated by the Kaplan-Meier method. RESULTS: In total, 804 patients with resectable G-NECs or G-MANECs were included, of whom 490 (60·9 per cent) received adjuvant chemotherapy. After propensity score matching, OS in the chemotherapy group was similar to that in the no-chemotherapy group. Among patients with G-NECs, survival in the fluorouracil (5-FU)-based chemotherapy group and the non-5-FU-based chemotherapy group was similar to that in the no-chemotherapy group. Similarly, etoposide plus cisplatin or irinotecan plus cisplatin was not associated with better OS in patients with G-NECs. Among patients with G-MANECs, OS in the non-5-FU-based chemotherapy group was worse than that in the no-chemotherapy group. Patients with G-MANECs did not have better OS when platinum-based chemotherapy was used. CONCLUSION: There was no survival benefit in patients who received adjuvant chemotherapy for G-NECs or G-MANECs.

ANTECEDENTES: El objetivo de este estudio fue evaluar si la quimioterapia adyuvante mejoraba la supervivencia en pacientes con carcinomas gástricos resecables neuroendocrinos (gastric neuroendocrine carcinomas, G-NECs) y carcinomas adenoneuroendocrinos mixtos (mixed adenoneuroendocrine carcinomas, G-MANECs). MÉTODOS: Se incluyeron pacientes con G-NECs y G-MANECs tratados quirúrgicamente en 21 centros en China entre 2004 y 2016. Se utilizó un análisis de emparejamiento por puntaje de propensión para reducir el sesgo de selección y el método de Kaplan-Meier para estimar la supervivencia global (overall survival, OS) de los pacientes en los diferentes grupos de tratamiento. RESULTADOS: En total, se incluyeron en el estudio 804 pacientes con G-NECs y G-MANECs resecables y 490 pacientes (60,9%) recibieron quimioterapia adyuvante. Después del emparejamiento por puntaje de propensión, la OS del grupo con quimioterapia fue similar a la del grupo sin quimioterapia. En los pacientes con G-NECs, la supervivencia en los grupos con quimioterapia basada en 5-FU (fluorouracilo) y de quimioterapia sin 5-FU fue similar a la del grupo sin quimioterapia. Asimismo, la combinación de etopósido y cisplatino o de irinotecán y cisplatino no se asoció con una mejor OS en pacientes con G-NECs. En pacientes con G-MANECs, la OS del grupo con quimioterapia sin 5-FU fue peor que la del grupo sin quimioterapia. Los pacientes con G-MANECs no presentaron una mejor OS cuando se administró quimioterapia basada en platinos. CONCLUSIÓN: La administración de quimioterapia adyuvante en pacientes con G-NECs y G-MANECs no mejoró la supervivencia.

MiR-30c exerts tumor suppressive functions in colorectal carcinoma by directly targeting BCL9.

OBJECTIVE: Colorectal carcinoma (CRC) remains a leading health threat worldwide due to its high mortality. MicroRNA (miR-30c) is an important tumor suppressor in various cancers. B cell lymphoma 9 (BCL9) is one of the candidate genes for cancers. The synergistic effects of miR-30c and BCL9 in CRC progression remain to be carefully elucidated. PATIENTS AND METHODS: Fifty pairs of CRC samples and matched adjacent non-tumor tissues were collected from Yantai Yuhuangding Hospital between 2015 and 2017. MiR-30c and BCL9 expression levels were measured by quantitative Real-time polymerase chain reaction (qRT-PCR) in CRC tissues and cell lines. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine the influence of miR-30c on the proliferation ability of CRC cells. Target Scan was used to predict the potential target of miR-30c. Then, luciferase assay was performed to confirm the prediction. In addition, we also investigated the biological influence of BCL9 on miR-30c-mediated functions in CRC. RESULTS: We found that miR-30c was significantly decreased in CRC tissues and cell lines while the BCL9 expression level was prominently increased in CRC tissues and cells. Additionally, the miR-30c expression was negatively correlated with BCL9 expressions in CRC tissues. Furthermore, the findings of this study also showed that BCL9 was a direct target of miR-30c in CRC and miR-30c could inhibit the CRC proliferation by binding to its 3'-UTR. CONCLUSIONS: This study showed that miR-30c overexpression inhibited CRC proliferation via the regulation of BCL9, suggesting that miR-30c may be a new molecular therapeutic target for CRC.

[The clinical significance of Delphian lymph node metastasis in papillary thyroid cancer].

Objective: To evaluate the clinical significance of Delphian lymph node (DLN) metastasis in papillary thyroid cancer (PTC). Method: A total of 505 cases with PTC confirmed pathologically in our hospital between January 2015 and December 2015 were retrospectively reviewed. 208 patients with DLN assessed separately by histopathologic examination who underwent primary surgery for PTC were included for the following analysis. Results: In 208 patients, the detection rate of DLN was 63.0% and the metastasis rate of DLN was 21.4%. DLN metastasis was correlated with PTC multifocality (P=0.038), tumor size over 1cm (P=0.001), BRAFV600E mutation (P=0.017) and central neck node metastasis (P<0.001). Tumor size over 1cm (95%CI 1.308-9.909, OR=3.600, P=0.013) and the number of node with central neck metastasis (95%CI 1.313-2.163, OR=1.685, P<0.001) were independent risk factors for DLN metastasis. The presence of DLN metastasis was associated with an 8.8-fold higher frequency of central neck node metastasis compared to cases without DLN metastasis. Among patients with DLN metastases, central lymph node metastasis was more common in the cases with lateral neck node metastases compared to those without lateral neck node metastases (6.5±3.0 vs 1.5±0.7, P=0.009), and 5 of the 6 patients also presented with PTC multifocality and BRAFV600E mutation. Conclusion: DLN metastasis implies a higher possibility of central neck lymph node metastasis. DLN should be assessed during operation to provide information for neck dissection, post-operative administration and follow-up strategy.

Characterization and comparison of transgenic Artemisia annua GYR and wild-type NON-GYR plants in an environmental release trial.

The anti-malarial drug, artemisinin, is quite expensive as a result of its slow content in Artemisia annua. Recent investigations have suggested that genetic engineering of A. annua is a promising approach to improve the yield of artemisinin. In this study, the transgenic A. annua strain GYR, which has high artemisinin content, was evaluated in an environmental release trial. First, GYR plants were compared with the wild-type variety NON-GYR, with regard to phenotypic characters (plant height, crown width, stem diameter, germination rate, leaf dry weight, 1000-seed weight, leave shape). Second, stress resistance in the two varieties (salt, drought, herbicide, and cold resistance) was evaluated under different experimental conditions. Finally, gene flow was estimated. The results indicated that there were significant differences in several agronomic traits (plant height, stem diameter, and leave dry weight) between the transgenic GYR and NON-GYR plants. Salt stress in transgenic and control plants was similar, except under high NaCl concentrations (1.6%, w/w). Leaf water, proline, and MDA content (increased significantly) were significantly different. Transgenic A. annua GYR plants did not grow better than NON-GYR plants with respect to drought and herbicide resistance. The two varieties maintained vitality through the winter. Third, gene flow was studied in an environmental risk trial for transgenic GYR. The maximum gene flow frequency was 2.5%, while the maximum gene flow distance was 24.4 m; gene flow was not detected at 29.2 m at any direction. Our findings may provide an opportunity for risk assessment in future commercialization of transgenic A. annua varieties.

[Percutaneous coronary intervention in eastern urban China: 2001-2011].

OBJECTIVE: We aimed to assess trends in clinical characteristics, treatment, and outcomes for hospitalized patients undergoing percutaneous coronary intervention (PCI) in eastern urban China from 2001 to 2011. METHOD: We analyzed a Chinese eastern representative sample of hospital admissions for PCI identified in China PEACE-retrospective CathPCI study using a two-stage random sampling design and calculated the weighted data of clinical information in each year. RESULTS: We included 3 308 admissions for PCI in 29 urban hospitals.Between 2001 and 2011, rates of hospitalizations for PCI increased by 15 fold.Compared with 2001, the patients undergoing PCI were more likely to be female, older than 70 years, and to have history of diabetes, hypertension, dyslipidemia and PCI in 2011.The proportion of trans radial PCIs was increased from 3.5% in 2001 to 72.6% in 2011 (Statistic=-28.95, Ptrend<0.000 1); the proportion of drug eluting stents (DES) among all the implanted stents was increased from 18.2%in 2001 to 98.4% in 2011(Statistic=-40.82, Ptrend<0.000 1), largely due to increased use of domestic DES.Less than 10% of medical records of patients undergoing primary PCI documented door time and balloon time.The median length of stay decreased from 13 days in 2001 to 10 days in 2011 (Statistic=-0.11, Ptrend<0.001). In-hospital mortality did not change significantly, but both any bleeding (Statistic= 2.66, Ptrend< 0.01) and access bleeding were decreased significantly (Statistic= 5.55, Ptrend< 0.000 1). CONCLUSIONS: During 2001 and 2011 in eastern urban China, there has been a rapid increase in the number and significant change in treatment patterns of PCI.Quality gaps are identified that represent opportunities to improve care.

[Progress in research of detection assay for pathogens causing community acquirerd pneumonia].

Community-acquired pneumonia(CAP)is a common respiratory infectious disease. The etiologic diagnosis of CAP remains an uneasy task. Early etiologic diagnosis is critical for proper treatment and might improve the prognosis. So, it is important to identify pathogens causing CAP in early time and accurate way with sensitive and effective method. This paper summarizes the recent progress in the research of the detection assay for CAP.

miR-218 tissue expression level is associated with aggressive progression of gastric cancer.

The aim of the present study was to investigate the clinical significance of microRNA-218 (miR-218) in gastric cancer. We enrolled 112 patients having undergone surgery for gastric cancer between May 2008 and June 2014. Expression of miR-218 was determined by real-time quantitative reverse transcription-polymerase chain reaction. Survival curves were plotted using the Kaplan-Meier method and compared by the log-rank test. We found that miR-218 expression was significantly downregulated in gastric cancer tissues compared to adjacent normal tissues (P < 0.001). Low miR-218 expression was significantly associated with tumor differentiation (P < 0.001), depth of tumor invasion (P = 0.006), and tumor node metastasis stage (P < 0.001). Kaplan-Meier survival analysis revealed that patients with low miR-218 levels showed significantly lower 5-year overall survival than those demonstrating high expression (P = 0.04). Multivariate Cox regression analyses indicated that low miR-218 expression constitutes an independent molecular biomarker for prediction of poor overall survival of gastric cancer patients (hazard ratio = 3.187, 95% confidence interval = 1.551-8.365, P = 0.037). In conclusion, miR-218 was remarkably downregulated in gastric cancer tissues and may serve as a prognostic biomarker for patients suffering from this disease.

[ST-segment elevation myocardial infarction in the eastern urban China: from 2001 to 2011].

OBJECTIVE: To assess trends in clinical characteristics, treatments, and outcomes for hospitalized patients with ST-segment elevation myocardial infarction(STEMI) in eastern urban China from 2001 to 2011. METHODS: The data were obtained from the China PEACE-retrospective acute myocardial infarction study. Patients admitted to hospital in the eastern urban China for STEMI were selected via two-stage random sampling. The first phase was to identify participating hospitals via a simple random-sampling procedure. The second stage was to select patients admitted to each participating hospitals for acute myocardial infarction in the year of 2001, 2006 and 2011 with a systematic sampling approach. Then clinical information was obtained via central medical record abstraction for each patient. In all analyses, weight was calculated proportional to the inverse sampling fraction for each period. Multilevel logistic regression models with generalized estimating equations were used for analysis of patient outcomes. RESULTS: This analysis included 5 257 patients with STEMI from 32 hospitals. In 2001, 2006, and 2011, the median age of STEMI patients was 66(57, 72)、67(56, 74)and 63(53, 74)years(trend test P=0.008), the proportion of female was 30.3%, 29.5% and 29.2%(trend test P=0.530), respectively. The proportion of cardiovascular risk factors increased over time(trend test P<0.001); 45.6%, 55.6%, and 56.3% patients had hypertension(trend test P<0.001); 18.8%, 27.7% and 26.2% patients had diabetes(trend test P<0.001); 50.1%, 59.2% and 70.5% patients had dyslipidemia(trend test P<0.001); 30.5%, 35.1% and 44.1% patients are current smokers(trend test P<0.001) in 2001, 2006 and 2011, respectively. Between 2001 and 2011, there were significant increases in aspirin use(80.7% in 2001, 90.4% in 2006, and 91.5% in 2011, trend test P<0.001), clopidogrel use(2.9% in 2001, 64.2% in 2006, and 90.3% in 2011, trend test P<0.001) within 24 hours after admission, statins use rate was 45.8% in 2001, 83.4% in 2006, and 93.8% in 2011(trend test P<0.001), and rate of direct percutaneous coronary intervention(PCI) was 21.0% in 2001, 29.7% in 2006, and 40.3% in 2011(trend test P<0.001) in patients without documented contraindications. However, the rate of reperfusion therapy was non-significantly decreased: 58.5% in 2001, 58.0% in 2006, and 55.5% in 2011 (trend test P=0.230). The use of beta blockers also decreased: 62.4% in 2001, 64.3% in 2006 and 55.2% in 2011(trend test P=0.001). The mortality rate within 7 days following admission was 7.8%, 7.0%, 6.1%, and the proportion of death or treatment withdrawal because of terminal status was 8.3%, 8.6%, 7.4% in 2001, 2006 and 2011, respectively. Both parameters were similar among the 3 time points(trend test P>0.05). CONCLUSIONS: During the past decade, there has been a rapid increase in application of new technology and drug for STEMI in the eastern urban China. However, important gaps persist between clinical practice and guideline recommendations, and the outcomes of patients have not been significantly improved. Clinical Trail Registry: ClinicalTrials.gov, NCT01624883.

Comparing the ability of luminex xMAP(®) salmonella serotyping assay and traditional serotyping method for serotyping salmonella isolated from southern Chinese population.

AIMS: To investigate whether the Luminex xMAP(®) Salmonella Serotyping Assay (xMAP SSA) is applicable to serotype Salmonella from humans in southern China. METHODS AND RESULTS: Two hundred and five Salmonella isolates from diarrhoea patients were serotyped by xMAP SSA in parallel with the traditional serotyping. Forty serotypes were identified among 205 isolates; the most prevalent serotypes identified were Salmonella Enteritidis, Salmonella Stanley, Salmonella I 4,5,12:i:-, and Salmonella Typhimurium. One hundred and ninety-five (95·1%, 195/205) isolates were serotyped completely by xMAP SSA, while 10 stereotypes were partially detected as they were not included in the assay. The xMAP SSA correctly identified 192 (98·5%, 192/195) isolates. Five nonmotile and three monophasic strains, which possessed flagellar antigen genes that weren't expressed, were completely serotyped by xMAP SSA; however, these isolates were left undetected by the traditional method. CONCLUSION: The xMAP SSA used in the study is a microsphere-based, molecular assay that could rapidly and accurately serotype Salmonella. It is suitable to identify the serovars of Salmonella in southern China. SIGNIFICANCE AND IMPACT OF THE STUDY: The xMAP SSA, with high-throughput characteristics, provides an accurate and rapid serotyping system that dramatically strengthens the capability of clinical and public health laboratories for Salmonella serotyping.

Screening of potential biomarkers for cholangiocarcinoma by integrated analysis of microarray data sets.

Cholangiocarcinoma (CCA) continues to harbor a difficult prognosis and it is difficult to diagnose in its early stages. The molecular mechanisms of CCA oncogenesis and progression are poorly understood. This study aimed to identify candidate biomarkers for CCA. Integrated analysis of microarray data sets was performed to identify differentially expressed genes (DEGs) between CCA and normal tissues. Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were then performed to identify the functions of DEGs. Furthermore, the protein-protein interaction (PPI) network of DEGs was constructed. The expressions of DEGs were validated in human CCA tissues by qRT-PCR. A set of 712 DEGs were identified in CCA compared with normal tissues, including 306 upregulated and 406 downregulated DEGs. It can be shown from the KEGG pathway analysis that some pathways may have important roles in pathology of CCA, including peroxisome proliferator-activated receptor signaling pathway, bile secretion, cell cycle, fat digestion and absorption. PPI network indicated that the significant hub proteins were PKM, SPP1 and TPM1. The abnormally overexpression PKM, SPP1 and TPM1 were closely related to oncogenesis and progression of CCA. PKM, SPP1, TPM1, COL1A1 and COL1A2 may serve as candidate biomarkers for diagnosis and prognosis of CCA.

Prevalence and clinical distribution of multidrug-resistant bacteria (3537 isolates) in a tertiary Chinese hospital (January 2012-December 2013).

OBJECTIVE: Multidrug-resistant organisms (MDROs) have become a widespread serious problem in recent years. Our objective was to determine the prevalence and clinical distribution of MDROs in a tertiary care hospital in China from January 1, 2012 to December 31, 2013. METHODS: The strains were cultured according to standard methods; bacterial identification and susceptibility testing were detected by Vitek 2 system. The prevalence and clinical distribution of extended-spectrum ß-lactamases (ESBLs)-producing enterobacteriaceae, carbapenem-resistant enterobacteriaceae (CRE), multiple-drug/pan-drug resistant P. aeruginosa (MDR/PDR-PA), carbapenem-resistant A. baumannii (CR-AB), methicillin-resistant S. aureus (MRSA) and vancomycin-resistant enterococcus (VRE) were analyzed by WHONET 5.6. RESULTS: A total of 3537 (33.4%) MDROs were found among 10,594 microbial isolates. ESBLs producing E. coli (ESBLs-ECO) (1153 cases) were the most frequent MDROs, followed by CR-AB (827 cases). The proportion of acquired resistance of A. baumannii (48.9%) accounted for the highest in all the MDROs. These MDROs were mainly isolated from respiratory (70.3%) and secretions (12.7%). Various types of intensive care unit (ICU) and surgery were the main source departments. The proportion of CRE and VRE were relatively few. CRE was most isolated from respiratory tract and closed body cavity fluid, while the distribution of VRE was relatively dispersed. CONCLUSION: High prevalence of MDROs has emerged in our hospital, particular in various ICU and surgical department. The effective way to prevent the further spread of MDROs is to strengthen the protection of respiratory tract and surgical wounds.

Event-specific qualitative and quantitative PCR detection of the GMO carnation (Dianthus caryophyllus) variety Moonlite based upon the 5'-transgene integration sequence.

To ensure the implementation of genetically modified organism (GMO)-labeling regulations, an event-specific detection method was developed based on the junction sequence of an exogenous integrant in the transgenic carnation variety Moonlite. The 5'-transgene integration sequence was isolated by thermal asymmetric interlaced PCR. Based upon the 5'-transgene integration sequence, the event-specific primers and TaqMan probe were designed to amplify the fragments, which spanned the exogenous DNA and carnation genomic DNA. Qualitative and quantitative PCR assays were developed employing the designed primers and probe. The detection limit of the qualitative PCR assay was 0.05% for Moonlite in 100 ng total carnation genomic DNA, corresponding to about 79 copies of the carnation haploid genome; the limit of detection and quantification of the quantitative PCR assay were estimated to be 38 and 190 copies of haploid carnation genomic DNA, respectively. Carnation samples with different contents of genetically modified components were quantified and the bias between the observed and true values of three samples were lower than the acceptance criterion (<25%) of the GMO detection method. These results indicated that these event-specific methods would be useful for the identification and quantification of the GMO carnation Moonlite.

Expression and hormonal regulation of E-cadherin in canine uterus during early pregnancy.

E-cadherin, a Ca(2+)-dependent cell adhesion molecule, is necessary for endometrial receptivity to blastocyst implantation. The aim of this study was to investigate the differential expression of E-cadherin in canine uterus during early pregnancy and its regulation under different conditions by in situ hybridization. E-cadherin mRNA expression was at a low level in the glandular epithelium on days 6, 12 and 17 of pregnancy. On days 20 and 23 of pregnancy, E-cadherin mRNA was highly expressed in the glandular epithelium surrounding the embryo, but not in the luminal epithelium and declined in villi and placenta on day 28 of pregnancy. During oestrous cycle, a moderate level of E-cadherin mRNA expression was found in the luminal and glandular epithelium of canine uteri at oestrus stage. The same expression was also found at anoestrus stage. Progesterone slightly induced the expression of E-cadherin mRNA in the luminal and glandular epithelium of ovariectomized canine uterus. These results suggest that E-cadherin expression is closely related to canine implantation and can be up-regulated by progesterone.

AAV9-mediated erythropoietin gene delivery into the brain protects nigral dopaminergic neurons in a rat model of Parkinson's disease.

We have recently shown that intrastriatal injection of recombinant human erythropoietin (EPO) protects dopaminergic (DA) neurons in the substantia nigra (SN) from 6-hydroxydopamine (6-OHDA) toxicity in a rat model of Parkinson's disease. However, systemic administration of EPO did not protect nigral DA neurons, suggesting that the blood-brain barrier limits the passage of EPO protein into the brain. In the present study, we used an adeno-associated viral (AAV) serotype 9 (AAV9) vector to deliver the human EPO gene into the brain of 6-OHDA-lesioned rats. We observed that expression of the human EPO gene was robust and stable in the striatum and the SN for up to 10 weeks. EPO-immunoreactive (IR) cells were widespread throughout the injected striatum, and EPO-IR neurons and fibers were also found in the ipsilateral SN. Enzyme-linked immunosorbent assay and western blot analyses exhibited dramatic levels of EPO protein in the injected striatum. As a result, nigral DA neurons were protected against 6-OHDA-induced toxicity. Amphetamine-induced rotational asymmetry and spontaneous forelimb use asymmetry were both attenuated. Interestingly, we also observed that intrastriatal injection of AAV9-EPO vectors led to increased numbers of red blood cells in peripheral blood. This highlights the importance of using an inducible gene delivery system for EPO gene delivery.

Addition of clopidogrel to aspirin in 45,852 patients with acute myocardial infarction: randomised placebo-controlled trial.

BACKGROUND: Despite improvements in the emergency treatment of myocardial infarction (MI), early mortality and morbidity remain high. The antiplatelet agent clopidogrel adds to the benefit of aspirin in acute coronary syndromes without ST-segment elevation, but its effects in patients with ST-elevation MI were unclear. METHODS: 45,852 patients admitted to 1250 hospitals within 24 h of suspected acute MI onset were randomly allocated clopidogrel 75 mg daily (n=22,961) or matching placebo (n=22,891) in addition to aspirin 162 mg daily. 93% had ST-segment elevation or bundle branch block, and 7% had ST-segment depression. Treatment was to continue until discharge or up to 4 weeks in hospital (mean 15 days in survivors) and 93% of patients completed it. The two prespecified co-primary outcomes were: (1) the composite of death, reinfarction, or stroke; and (2) death from any cause during the scheduled treatment period. Comparisons were by intention to treat, and used the log-rank method. This trial is registered with ClinicalTrials.gov, number NCT00222573. FINDINGS: Allocation to clopidogrel produced a highly significant 9% (95% CI 3-14) proportional reduction in death, reinfarction, or stroke (2121 [9.2%] clopidogrel vs 2310 [10.1%] placebo; p=0.002), corresponding to nine (SE 3) fewer events per 1000 patients treated for about 2 weeks. There was also a significant 7% (1-13) proportional reduction in any death (1726 [7.5%] vs 1845 [8.1%]; p=0.03). These effects on death, reinfarction, and stroke seemed consistent across a wide range of patients and independent of other treatments being used. Considering all fatal, transfused, or cerebral bleeds together, no significant excess risk was noted with clopidogrel, either overall (134 [0.58%] vs 125 [0.55%]; p=0.59), or in patients aged older than 70 years or in those given fibrinolytic therapy. INTERPRETATION: In a wide range of patients with acute MI, adding clopidogrel 75 mg daily to aspirin and other standard treatments (such as fibrinolytic therapy) safely reduces mortality and major vascular events in hospital, and should be considered routinely.

Early intravenous then oral metoprolol in 45,852 patients with acute myocardial infarction: randomised placebo-controlled trial.

BACKGROUND: Despite previous randomised trials of early beta-blocker therapy in the emergency treatment of myocardial infarction (MI), uncertainty has persisted about the value of adding it to current standard interventions (eg, aspirin and fibrinolytic therapy), and the balance of potential benefits and hazards is still unclear in high-risk patients. METHODS: 45,852 patients admitted to 1250 hospitals within 24 h of suspected acute MI onset were randomly allocated metoprolol (up to 15 mg intravenous then 200 mg oral daily; n=22,929) or matching placebo (n=22,923). 93% had ST-segment elevation or bundle branch block, and 7% had ST-segment depression. Treatment was to continue until discharge or up to 4 weeks in hospital (mean 15 days in survivors) and 89% completed it. The two prespecified co-primary outcomes were: (1) composite of death, reinfarction, or cardiac arrest; and (2) death from any cause during the scheduled treatment period. Comparisons were by intention to treat, and used the log-rank method. This study is registered with ClinicalTrials.gov, number NCT 00222573. FINDINGS: Neither of the co-primary outcomes was significantly reduced by allocation to metoprolol. For death, reinfarction, or cardiac arrest, 2166 (9.4%) patients allocated metoprolol had at least one such event compared with 2261 (9.9%) allocated placebo (odds ratio [OR] 0.96, 95% CI 0.90-1.01; p=0.1). For death alone, there were 1774 (7.7%) deaths in the metoprolol group versus 1797 (7.8%) in the placebo group (OR 0.99, 0.92-1.05; p=0.69). Allocation to metoprolol was associated with five fewer people having reinfarction (464 [2.0%] metoprolol vs 568 [2.5%] placebo; OR 0.82, 0.72-0.92; p=0.001) and five fewer having ventricular fibrillation (581 [2.5%] vs 698 [3.0%]; OR 0.83, 0.75-0.93; p=0.001) per 1000 treated. Overall, these reductions were counterbalanced by 11 more per 1000 developing cardiogenic shock (1141 [5.0%] vs 885 [3.9%]; OR 1.30, 1.19-1.41; p<0.00001). This excess of cardiogenic shock was mainly during days 0-1 after admission, whereas the reductions in reinfarction and ventricular fibrillation emerged more gradually. Consequently, the overall effect on death, reinfarction, arrest, or shock was significantly adverse during days 0-1 and significantly beneficial thereafter. There was substantial net hazard in haemodynamically unstable patients, and moderate net benefit in those who were relatively stable (particularly after days 0-1). INTERPRETATION: The use of early beta-blocker therapy in acute MI reduces the risks of reinfarction and ventricular fibrillation, but increases the risk of cardiogenic shock, especially during the first day or so after admission. Consequently, it might generally be prudent to consider starting beta-blocker therapy in hospital only when the haemodynamic condition after MI has stabilised.

[Medical application of hyaluronic acid and its derivatives].

OBJECTIVE: To review the recent advances of hyaluronic acid and its derivatives in medical application. METHODS: Recent original articles related to hyaluronan derivatives and their medical applications were retrieved extensively. RESULTS: Hyaluronic acid and its derivatives play important roles in visosurgery, arthritis therapy, prevention of adhesion, drug delivery, soft-tissue dilation, and percutaneous embolization. CONCLUSION: Development of hyaluronan derivatives may widen their medical application.

Chromosome breakage at a major fragile site associated with P-glycoprotein gene amplification in multidrug-resistant CHO cells.

Recent studies of several drug-resistant Chinese hamster cell lines suggested that a breakage-fusion-bridge mechanism is frequently involved in the amplification of drug resistance genes. These observations underscore the importance of chromosome breakage in the initiation of DNA amplification in mammalian cells. However, the mechanism of this breakage is unknown. Here, we propose that the site of chromosome breakage consistent with the initial event of P-glycoprotein (P-gp) gene amplification via the breakage-fusion-bridge cycle in three independently established multidrug-resistant CHO cells was located at 1q31. This site is a major chromosome fragile site that can be induced by methotrexate and aphidicolin treatments. Pretreatments of CHO cells with methotrexate or aphidicolin enhanced the frequencies of resistance to vinca alkaloid and amplification of the P-gp gene. These observations suggest that chromosome fragile sites play a pivotal role in DNA amplification in mammalian cells. Our data are also consistent with the hypothesis that gene amplification can be initiated by stress-induced chromosome breakage that is independent of modes of action of cytotoxic agents. Drug-resistant variants may arise by their growth advantage due to overproduction of cellular target molecules via gene amplification.