Molecular and immunological characteristics of patients with CMTM6 low expression colorectal cancer.

CKLF-like MARVEL transmembrane domain-containing 6 (CMTM6), a regulator of programmed cell death ligand 1 (PD-L1), has attracted extensive attention due to its role in tumors. However, research on the expression of CMTM6 in colorectal cancer (CRC) and its relationship with PD-L1 expression and immune cell infiltration is limited. We used The Cancer Genome Atlas database to mine and analyze data from patients with CRC using bioinformatics methods. We investigated the expression of CMTM6 in CRC and its relationship with PD-L1 expression and immune cell infiltration. Immunohistochemistry and PCR were performed to detect CMTM6 and PD-L1 expression in CRC tissues. Differential gene expression analysis was performed using the edgeR package in R and immune cell infiltration analysis was performed using the ssGSEA algorithm. Additionally, GO and KEGG enrichment analyses were conducted to identify the biological processes and pathways associated with low CMTM6 expression. Our study found that CMTM6 expression was significantly upregulated in CRC tissues compared to that in adjacent normal tissues. Patients with high CMTM6 expression exhibited significantly increased levels of PD-L1 expression and higher levels of tumor-infiltrating immune cells compared to patients with low CMTM6 expression. GO and KEGG analyses suggested that CMTM6 may be involved in multiple immune regulatory pathways in CRC.

[Determination of kojic acid in fermented foods by solid-phase extraction coupled with ultra performance liquid chromatography-tandem mass spectrometry].

Kojic acid naturally appears in fermented foods and can be formed during the aerobic fermentation process induced by Aspergillus and Penicillium fungi. It is widely used in the food industry because it exhibits antibacterial and antifungal properties and does not affect food taste. However, recent studies indicate that kojic acid may be a potential carcinogen. Therefore, assessing the health risks of kojic acid in fermented foods are of great importance, and developing a sensitive and accurate analytical method for this compound is a significant endeavor. Much efforts have been devoted to the detection of kojic acid using electrochemistry, high performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). HPLC and HPLC-MS/MS are the analytical techniques most often employed for this purpose. Of these two methods, HPLC-MS/MS displays excellent sensitivity and is the optimal selective technique. Pretreatment is usually necessary for kojic acid determination because of the complex matrix effects of fermented foods. However, few researches on the determination of kojic acid in food are available, and, to the best of our knowledge, the determination of kojic acid using solid-phase extraction (SPE) pretreatment has not been reported yet. Herein, a convenient, sensitive, and accurate method was developed to determine kojic acid in fermented foods using solid-phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS). The pretreatment conditions, such as the extraction solvent, cartridge, rinse solvent, and eluent, were systematically optimized. The samples, including soy sauce, vinegar, liquor, sauce, fermented soya bean, and fermented bean curd, were extracted with 0.1% formic acid-absolute ethyl alcohol and purified using a PRiME HLB cartridge. Kojic acid was separated using an ACQUITY UPLC® BEH C18 column (100 mm×2.1 mm, 1.7 µm) with formic acid-acetonitrile (1∶999, v/v) and formic acid-5 mmol/L ammonium acetate (1∶999, v/v) solutions as the mobile phases under gradient elution mode. MS was performed in electrospray positive ionization (ESI+) and multiple reaction monitoring (MRM) modes. An internal standard method was used for quantification. Under optimized conditions, good linearity was achieved at mass concentrations of 5.0-100.0 µg/L, with a correlation coefficient (r) of 0.9994. The limits of detection and quantification of the method for kojic acid were 2-5 µg/kg and 6-15 µg/kg, respectively. Good recoveries of 86.8%-111.7%, intra-day precisions of 1.0%-7.9% (n=6), and inter-day precisions of 2.7%-10.2% (n=5) were also obtained. The matrix effect was evaluated by establishing a matrix-matching calibration curve, and weak inhibitory effects were found in vinegar and liquor; moderate inhibitory effects in fermented bean curd, fermented soya bean, and soy sauce; and a strong inhibitory effect in sauce. The developed method was used to detect kojic acid in 240 fermented foods, and the results showed that the detection rate of vinegar was the highest, followed by liquor, sauce, soy sauce, fermented soya bean, and fermented bean curd, the contents were 5.69-2272 µg/kg. Matrix interferences can be significantly reduced by optimizing the pretreatment and detection procedures. The proposed method is sensitive, accurate, and can be used to analyze kojic acid in fermented foods.

[Rapid determination of five halobenzoquinones in aquatic products by QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry].

Halobenzoquinones (HBQs), which are emerging chlorinated disinfection byproducts (DBPs), have attracted increasing attention because they are frequently detected in treated tap water, entrainment water, etc. These compounds are mainly generated during the water treatment process using chlorine, chloramine, and chlorine dioxide as disinfectants, and display more toxic effects than regulated DBPs, such as trihalomethane and haloacetic acid. HBQs have been recognized as potential bladder carcinogens and are harmful to the nervous system. Additionally, they can exert genotoxic effects and cause oxidative damage to DNA and proteins. The risk of HBQs in aquatic products is expected to rise because the disinfection of public facilities has significantly increased in recent years. Therefore, developing a sensitive and accurate analytical method to detect HBQs in aquatic products is of great importance. Several analytical methods, including gas chromatography, gas chromatography-mass spectrometry, electrochemical methods, liquid chromatography, and liquid chromatography-tandem mass spectrometry, can be used to identify and quantify HBQs in water. However, to the best of our knowledge, no reports on the determination of HBQ levels in aquatic products are yet available. Further, pretreatment is essential for HBQ determination because of the complex matrix effects of aquatic products. Herein, a sensitive and accurate method based on the QuEChERS technique coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the simultaneous determination of five HBQs in aquatic products. For the QuEChERS procedure, the pretreatment conditions, such as the extraction solvent and adsorbent species, were systematically optimized. The sample was extracted with 10 mL of 10% methanol acetonitrile solution (containing 0.1% formic acid), dehydrated, and centrifuged with sodium chloride and anhydrous magnesium sulfate. The supernatant was purified using a QuEChERS packing material consisting of 50 mg N-propylethylenediamine (PSA), 30 mg of graphitized carbon black (GCB), and 30 mg of neutral alumina (Al2O3), dried with nitrogen, and concentrated. The five HBQs were separated on a Waters ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 µm) using 0.25% acetonitrile formate solution and 0.25% formic acid aqueous solution as the mobile phase under a gradient elution program and then detected using UPLC-MS/MS with negative electrospray ionization (ESI-) under multiple reaction monitoring (MRM) mode. Quantitative analysis was performed using a matrix-matched external standard method. The five HBQs achieved rapid separation within 6 min, indicating that the proposed method has a much shorter separation time compared with previous studies. The matrix effect was evaluated by establishing a matrix-matched calibration curve. The results showed that 2,5-dichloro-1,4-benzoquinone (2,5-DCBQ) presented a matrix-enhancing effect, whereas the other HBQs displayed matrix-inhibiting effects. In particular, tetrachlorobenzoquinone (TCBQ) exhibited strong inhibitory effects. Under the optimized experimental conditions, the five HBQs demonstrated good linear relationships in the range of 1.0-50.0 µg/L, with correlation coefficients (r)≥0.9992. The detection limits of the method were 0.15-0.8 µg/kg, and the recoveries of the target compounds were 85.9%-116.5%. The relative standard deviations were 1.4%-8.2%, which indicates good reproducibility. The proposed method was successfully applied to actual sample detection, and 2,6-dichloro-3-methyl-1,4-benzoquinone (2,6-DCMBQ) was detected in grass carp. The proposed method is convenient, sensitive, accurate, and suitable for the simultaneous determination of five HBQs in aquatic products. Moreover, the developed method provides a reliable reference for the routine monitoring of trace HBQs in food samples.

Determination of the Dose-Response Relationship of Epidural Dexmedetomidine Combined with Ropivacaine for Labor Analgesia.

Background: The safety and efficacy of dexmedetomidine for epidural labor analgesia have been reported in numerous literatures, but the optimal dose has not been fully determined. The objective of this study was to determine the dose-response relationship of epidural dexmedetomidine (combined with ropivacaine) for labor analgesia. Methods: A total of 120 full-term laboring parturients requesting epidural labor analgesia were enrolled in the study from July 5, 2020 to September 22, 2021. The parturients were randomly assigned to receive 0, 0.1, 0.2, 0.3, 0.4 or 0.5 µg/mL dexmedetomidine combined with 0.075% ropivacaine epidurally. An effective dose was defined as numerical rating scale (NRS) pain score ≤3 at 30-minutes of epidural drug injection. The dose-response relationship of dexmedetomidine (with ropivacaine) for epidural labor analgesia was performed using probit regression. The median effective dose (ED50) and the 95% effective dose (ED95) values for epidural dexmedetomidine combined with 0.075% ropivacaine with 95% confidence intervals (CIs) were derived by interpolation. Results: The estimated values of ED50 and ED95 with 95% CIs for epidural dexmedetomidine (combined with 0.075% ropivacaine) were 0.085 (0.015 to 0.133) µg/mL and 0.357 (0.287 to 0.493) µg/mL, respectively. No differences were found among groups for sensory block level, number of parturients with Bromage score >0, total dosage of analgesics, cesarean delivery rate, fetal birth weight, Apgar score at 1-minute, Apgar score at 5-minutes and adverse effects. Compared with other groups, group dexmedetomidine 0.5 µg/mL had a longer duration of the first stage of labor. Conclusion: The ED50 and ED95 values of dexmedetomidine for epidural labor analgesia was 0.085 and 0.357 µg/mL under the conditions of this study. Dexmedetomidine is a suitable adjuvant for epidural labor analgesia.

Statin use and survival among women with ovarian cancer: an Australian national data-linkage study.

BACKGROUND: Five-year ovarian cancer survival rates are below 50%; there is considerable interest in whether common medications like statins may improve survival. METHODS: We identified women diagnosed with ovarian cancer in Australia from 2003 to 2013 through the Australian Cancer Database and linked these records to national medication and death databases. We used Cox proportional hazards regression to estimate hazard ratios (HR) and confidence intervals (CI) for associations between statins and survival. RESULTS: Pre-diagnosis statin use was not associated with survival overall but was associated with better survival among women with endometrioid cancers. Statin use after diagnosis was associated with better ovarian cancer-specific survival (OVS, HR = 0.87, 95%CI 0.81-0.94), but this association was largely restricted to women who started using statins after their cancer diagnosis (OVS HR = 0.68, 0.57-0.81 vs. HR = 0.94, 0.87-1.01 for continuing users). The association was strongest for endometrioid cancers (OVS HR = 0.48, 0.29-0.77). CONCLUSIONS: Use of statins may confer a survival benefit for women with ovarian cancer but it is impossible to rule out bias in observational studies. Particularly problematic are reverse causation where disease status affects statin use, confounding by indication and the absence of data for women with normal cholesterol levels. A randomised trial is required to provide definitive evidence.

Is there sufficient evidence to recommend women diagnosed with endometrial cancer take a statin: Results from an Australian record-linkage study.

OBJECTIVE: A recent paper suggested all women with endometrial cancer should take statins but it is unclear whether there is sufficient evidence to justify this recommendation. METHODS: We identified all women diagnosed with uterine cancer in Australia between July 2003 and December 2013 (2012 in New South Wales) through the Australian Cancer Database (N = 16,501) and linked these to the national prescription database and National Death Index to identify statin use and survival outcomes to December 2015. We used Cox proportional hazards regression to estimate hazard ratios (HR) and 95% confidence intervals (CI) for the associations between statin use and survival. RESULTS: Among the 15,703 women with endometrial cancer, pre-diagnosis statin use was not associated with survival. Endometrial cancer-specific mortality was lower among women who used statins after diagnosis (time-varying models: HR = 0.92; 95%CI 0.82-1.03) but the association was only seen among women with type 1 cancers (0.87; 0.76-1.00), for hydrophilic statins (0.84; 0.68-1.03) and for new use of statins after diagnosis (0.75; 0.59-0.95). There was a weak dose-response with increasing number of statin prescriptions. Sensitivity analyses using inverse probability of treatment weights were similar. CONCLUSION: Women with endometrial cancer who take statins after diagnosis may have better survival than those who do not use statins. However, it is impossible to completely rule out bias, particularly reverse causation where disease status may affect statin use. We believe it is too early to recommend all women with endometrial cancer take statins, but there is sufficient evidence to justify a randomized trial.

Metformin and cancer-specific survival among breast, colorectal, or endometrial cancer patients: A nationwide data linkage study.

AIMS: Equivocal results of association between metformin and cancer-specific survival need more investigation. We tested the hypothesis that adherence to the drug had a lower cancer-specific mortality in a homogeneous population (i.e. regular users). METHODS: The Australian Cancer database was linked to the Pharmaceutical Benefits Scheme data and the National Death Index. Adherence to metformin was calculated by proportion of days covered. Cox regression models with time-varying covariates were used to estimate multivariable-adjusted cause-specific hazard ratios (HRs) and 95% confidence intervals (CI) for the association of adherence to metformin and cancer-specific mortality. RESULTS: Between 2003 and 2013, three separate cohorts of 6717, 3121, and 1854 female patients were identified with newly diagnosed breast, colorectal, or endometrial cancer. The 1-year adherence was similar at baseline in three cohorts, on average 75%. Each 10% increase in 1-year adherence to metformin reduced cancer-specific mortality among women with breast cancer (adjusted HR = 0.95; 95% CI, 0.93-0.97), colorectal cancer (adjusted HR = 0.94; 95% CI, 0.91-0.96), or endometrial cancer (adjusted HR = 0.95; 95% CI, 0.90-0.99). The inverse associations remained unchanged in most subgroup analyses. CONCLUSIONS: Among metformin users, adherence to this drug is inversely associated with reduced cancer-specific mortality. If confirmed, metformin could be considered as an adjuvant treatment.

Association between evidence-based medication at discharge and outcomes in patients with heart failure: a systematic review and meta-analysis.

To conduct a meta-analysis of observational studies assessing the association between dispensing evidence-based medications (EBMs) at discharge and outcomes, we extracted published studies in English from PubMed, Medline, and EMBASE from 2007 to early 2019. The EBMs included renin-angiotensin system inhibitors (RASIs), ß-blockers, and mineralocorticoid receptor antagonists (MRAs). The main outcomes of interest were all-cause death and heart failure (HF) readmission. Pooled hazard ratios (HRs) were calculated using random effect model from the adjusted HRs or relative risks (RRs) extracted from individual studies, stratified by HF patients with reduced ejection fraction (HFrEF), and preserved ejection fraction (HFpEF). Forty-three studies including a total number of 295,060 patients with an average follow-up time of 2.3 years were identified for systematic review. Dispensing RASI at discharge was independently associated with 30% and 25% lower risks of all-cause death and HF readmission respectively in HFrEF but has a moderate effect on reducing all-cause deaths (HR = 0.88, 95% CI: 0.81-0.95) in HFpEF. By contrast, dispensing ß-blockers at discharge was associated with 35% lower risk of all-cause deaths in HFrEF and has a weak association with borderline statistical significance on improving overall survival in HFpEF. Dispensing MRA at discharge was associated with 5% lower risk of all-cause death in HFrEF. This meta-analysis provides evidence to support RASIs and ß-blockers as primary pharmacotherapies for HF patients. Our findings suggest that the health professionals maintain use of RASIs and ß-blockers at discharge for potential survival improvement.

Does adherence to lipid-lowering medications improve cancer survival? A nationwide study of breast and colorectal cancer, and melanoma.

AIMS: Inconclusive findings of lipid-lowering medications (LLMs) on cancer survival benefit require more evidence. We tested the hypothesis that adherence to this drug is associated with reduced cancer-specific mortality in a homogeneous population who had used this drug before cancer diagnosis. METHODS: The Australian Cancer Database was linked to the Pharmaceutical Benefits Scheme database, and to the National Death Index (up to 2015). Medication adherence was calculated by proportion of days covered. Cox regression models with time-varying covariates were used to derive multivariable-adjusted cause-specific hazard ratio (HR) and 95% confidence interval (CI) for the associations between adherence to LLMs, statins, lipophilic, and hydrophilic statins and cancer-specific mortality. RESULTS: From 2003 to 2013, 3 separate cohorts of 20 046, 11 719 and 6430 female patients with newly diagnosed breast, colorectal cancer, and melanoma respectively were identified. The 1-year adherence was similar at 1-year prediagnosis in the 3 cohorts, on average 82%. Each 10% increase in 1-year adherence to LLMs was inversely associated with cancer-specific mortality among women with breast cancer (fully adjusted HR = 0.92, 95% CI 0.91-0.93), colorectal cancer (fully adjusted HR = 0.92, 95% CI 0.91-0.93), or melanoma (fully adjusted HR = 0.97, 95% CI 0.94-1.00). The reductions in cancer-specific mortality were more pronounced for women who adhered to lipophilic than hydrophilic statins in all 3 cancers albeit not statistically significant for melanoma. CONCLUSION: Among LLM users, adherence to this drug is associated with a decrease in cancer-specific mortality. If confirmed, LLMs could be considered as an adjuvant cancer therapy to improve prognosis in cancer survivors.

Lipid-lowering medication use and cancer-specific survival among endometrial or lung cancer patients: an Australian nationwide cohort study.

PURPOSE: Inconsistent results of lipid-lowering medications (LLMs) on improved cancer survival need more investigations. We tested the hypothesis that adherence to the drug would be associated with a lower cancer-specific mortality in a homogeneous population who has ever used the drug. METHODS: Utilising data from the Australian Cancer database, linked to the Pharmaceutical Benefits Scheme data and the National Death Index, we identified two separate cohorts of 4519 and 3083 women patients with newly diagnosed endometrial and lung cancer respectively between 2003 and 2013. Adherence to this drug was calculated by proportion of days covered. Cox regression models with time-varying covariates were used to estimate the multivariable-adjusted cause-specific hazard ratios (HRs) and 95% confidence intervals (CIs) for the association of adherence to LLMs, statins, lipophilic and hydrophilic statins, and cancer-specific mortality. RESULTS: Each 10% increase in 1-year adherence to LLMs reduced cancer-specific mortality among women with endometrial cancer (adjusted HR=0.93, 95% CI 0.90-0.96) or lung cancer (adjusted HR=0.95, 95% CI 0.93-0.97). The inverse associations remained unchanged in different subgroup analyses. The reductions in lung cancer mortality were not apparent for women who adhered to lipophilic statins albeit better endometrial cancer survival appeared in the lipophilic statin group and borderline statistical improvement in the hydrophilic statin group. CONCLUSIONS: Among LLM users, adherence to this drug is inversely associated with reduced cancer-specific mortality. Together with previous evidence, randomised controlled trials are called for to confirm whether LLMs could be considered as an adjuvant treatment to improve prognosis.

Incidence and Predictors of Sudden Cardiac Death After a Major Non-Fatal Cardiovascular Event.

BACKGROUND: Sudden cardiac death (SCD) still accounts for the majority of deaths from the four major cardiovascular events (myocardial infarction (MI), heart failure (HF), atrial fibrillation (AF), and stroke) despite substantial progress on prevention. METHODS: Four separate cohorts (one for each of the four major cardiovascular conditions) were captured through person-linked hospital morbidity and mortality data collections between 2000 and 2009 and followed-up for 11.5 years. The incidence rate for each cohort was total SCD cases divided by sum of follow-up time for each individual alive. Kaplan-Meier survival curve was used to calculate unadjusted risk of SCD. Predictors of SCD were identified by fitting multivariable adjusted Cox regression models in each of the cohorts. RESULTS: There were 1,174 cases of SCD from 53,614 total CVD events across the cohorts (35.6% for MI, 15.6% for HF, 22.4% for AF, 26.4% for stroke). The incidence rate and unadjusted risk of SCD were both highest after incident hospitalisation for HF, followed by MI, stroke and AF. The elevated risk of SCD was independently associated with MI, HF, arrhythmias, peripheral artery disease, diabetes, chronic kidney disease, and prior coronary heart disease (hazard ratios ranging from 1.1 to 2.8). Early revascularisation is protective in 28-day survivors after an incident MI event. CONCLUSIONS: An appreciable incidence of SCD following an incident event of MI, HF, AF and stroke deserves greater prevention efforts. Major medical conditions such as MI, HF, peripheral artery disease, and arrhythmias are risk markers of SCD and coronary revascularisation is protective.

Combined effects of aspirin and vitamin D3 on two OSCC cell lines: a preliminary study.

OBJECTIVES: We evaluated the potential effects of aspirin combined with vitamin D3 on cell proliferation and apoptosis in oral cancer cells. RESULTS: Compared to the untreated control or individual drug, the combinations of aspirin and vitamin D3 significantly decreased the rates of cell proliferation by CCK-8 assay, and caused higher rates of cell apoptosis in both CAL-27 and SCC-15 cells by Annexin V-FITC apoptosis assay and flow cytometry. Remarkably, the combined treatment with aspirin and vitamin D3 significantly suppressed the expression of Bcl-2 protein and p-Erk1/2 protein, examined by western blot analysis. CONCLUSIONS: Our study demonstrates that aspirin and vitamin D3 have biological activity against two human OSCC cell lines and their activity is synergistic or additive when two drugs used in combination with therapeutic concentrations. The combination of aspirin and vitamin D3 may be an effective approach for inducing cell death in OSCC.

Research on the Determination of 10 Industrial Dyes in Foodstuffs.

An efficient method was developed for the simultaneous determination of 10 industrial dyes (basic orange 2, basic orange 21, basic orange 22, acid orange II, auramine, basic rhodamine B and Sudan I-IV) in the foodstuffs using high-performance liquid chromatography coupled with diode array detector. Samples were extracted with acetonitrile and cleaned up on a solid phase extraction cartridge using HLB. The chromatographic separation was achieved on a C18 column using a mobile phase consisting of methanol and 10 mmol/L ammonium acetate with 0.1% formic acid by gradient elution. Good linearity (r ≥ 0.9993) was observed between 0.050 and 5.0 µg/mL. The limits of detection were in the range of 0.007-0.01 mg/kg, high recoveries (80.6-104%) and good reproducibility (1.1-5.7%) were obtained. Such method is simple, feasible and accurate, which can be applied to the quantification of 10 dyes in food samples.

Temporal Trends in Sudden Cardiac Death From 1997 to 2010: A Data Linkage Study.

BACKGROUND: Community-wide trends data for sudden cardiac death (SCD) are scarce, unlike widely reported declines in cardiovascular disease (CVD) mortality. Using administrative data, we aimed to examine population-level trends in SCD, stratified by sex, age and prior CVD hospitalisation. METHODS: Person-linked mortality and hospital morbidity data were used to identify SCD and determine hospitalisation and comorbidity using a 10-year hospitalisation lookback period. Log-linear Poisson regression was used to calculate annual rate changes and rate ratios. RESULTS: In Western Australia, 7160 SCD cases were identified from 1997 to 2010 with males comprising 69%. Overall age-standardised SCD rates decreased by 17% in men and 31% in women from 1997-2001 to 2007-2010. The annual rate reduction was higher in women than men (-4.0%/year versus -2.3%/year; p=0.0039). Significant reductions were observed for 55-69 year-old and 70-84 year-old men and women but not for the 35-54 year-olds. The overall relative risk comparing men to women increased slightly from 2.4 in 1997 to 3.0 in 2010 (trend p=0.0039) but differed across age groups. The relative risk declined in 35-54 year-olds from 5.1 to 3.2 whereas it increased from 2.9 to 3.9 in 55-69 year-olds and 1.9 to 2.3 in 70-84 year-olds. Declining trends in SCD rates were observed in those with and without prior CVD and were similar to CVD mortality trends (-4.9%/year in men and -5.5%/year in women). CONCLUSIONS: Trends in rates of SCD fell in middle to older aged men and women, with and without CVD, and mirrored the fall in fatal CVD. Limited improvement in 35-54 year-olds requires further investigation.

Sudden cardiac death rates in an Australian population: a data linkage study.

OBJECTIVE: The aim of the present study was to develop criteria to identify sudden cardiac death (SCD) and estimate population rates of SCD using administrative mortality and hospital morbidity records in Western Australia. METHODS: Four criteria were developed using place, death within 24 h, principal and secondary diagnoses, underlying and associated cause of death, and/or occurrence of a post mortem to identify SCD. Average crude, age-standardised and age-specific rates of SCD were estimated using population person-linked administrative data. RESULTS: In all, 9567 probable SCDs were identified between 1997 and 2010, with one-third aged ≥ 35 years having no prior admission for cardiovascular disease. SCD was more frequent in men (62.1%). The estimated average annual crude SCD rate for the period was 34.6 per 100 000 person-years with an average annual age-standardised rate of 37.8 per 100 000 person-years. Age-specific standardised rates were 1.1 per 100 000 person-years and 70.7 per 100 000 person-years in people aged 1-34 and ≥ 35 years, respectively. Ischaemic heart disease (IHD) was recorded as the underlying cause of death in approximately 80% of patients aged ≥ 35 years, followed by valvular heart disease and heart failure. IHD was the most common cause of death in those aged 1-34 years, followed by unspecified cardiomyopathy and dysrhythmias. CONCLUSIONS: Administrative morbidity and mortality data can be used to estimate rates of SCD and therefore provide a suitable methodology for monitoring SCD over time. The findings highlight the magnitude of SCD and its potential for public health prevention.

Development of an ERIC sequence typing scheme for Laribacter hongkongensis, an emerging pathogen associated with community-acquired gastroenteritis and travellers' diarrhoea.

Laribacter hongkongensis is a potential emerging pathogen, associated with community-acquired diarrhoea. For epidemiological purposes, different molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing, have been developed for this pathogen. However, these methods require specialized equipment and costly reagents. More importantly, they are labour-intensive and time-consuming, which is not really suitable for foodborne disease outbreak investigations. In this study, we developed a rapid and reliable method using 22-mer primers specific for the enterobacterial repetitive intergenic consensus sequence (ERIC). PFGE was used for comparison, to evaluate this method. A total of 81 isolates of L. hongkongensis were examined: 79 isolates recovered from food of diverse origins and two strains derived from patients with L. hongkongensis-associated infection. Typing patterns and clustering analysis indicated that the 81 L. hongkongensis isolates were grouped into 21 and 13 genotypes by ERIC-PCR and PFGE, respectively. ERIC-PCR was found as reproducible as PFGE. A high percentage (70.4%) of isolates yielded distinguishable ERIC-PCR patterns, which were concordant with the results from PFGE. These results suggest that ERIC-PCR is valuable for use in the epidemiological investigation of L. hongkongensis.

The prevalence, antimicrobial resistance and PFGE profiles of Laribacter hongkongensis in retail freshwater fish and edible frogs of southern China.

Laribacter hongkongensis is a novel emerging pathogen associated with human gastroenteritis. We aimed to investigate the prevalence, antimicrobial resistance and genotypic relationship of 199 L. hongkongensis isolates from 690 intestinal samples of fish and frogs. These samples were collected from retail markets in the city of Guangzhou in southern China from October 2008 to September 2009. L. hongkongensis was detected in from 80 (16.3%) out of 490 freshwater fish, and this number included 76 (32.3%) out of 235 grass carp and 4 (14.8%) out of 27 bighead carp. A higher isolation rate of 59.5% (119 out of 200) was observed in edible frogs. The isolation rate was highest in the spring in comparison with other seasons. Notably, 63.8% of the isolates were resistant to at least one class of antimicrobial agents. Analysis by pulsed-field gel electrophoresis (PFGE) revealed that the isolates could be grouped into three clusters. Isolates from fish intestines were grouped into two clusters: cluster I and II. Isolates of frog-origin and several fish-origin isolates were grouped into cluster III. Two patient-derived strains could be classed into cluster III. Extensive genetic heterogeneity among the isolates was observed. The results indicate that L. hongkongensis isolates exhibits host tropism, extensive resistance to widely used antimicrobials and diverse biological evolution in an aquatic environment. The frog is more likely than the freshwater fish to be the potential source for human infection with L. hongkongensis.

Induction of a high-yield lovastatin mutant of Aspergillus terreus by ¹²C6⁺ heavy-ion beam irradiation and the influence of culture conditions on lovastatin production under submerged fermentation.

Heavy-ion beams, possessing a wide mutation spectrum and increased mutation frequency, have been used effectively as a breeding method. In this study, the heavy-ion beams generated by the Heavy-Ion Research Facility in Lanzhou were used to mutagenize Aspergillus terreus CA99 for screening high-yield lovastatin strains. Furthermore, the main growth conditions as well as the influences of carbon and nitrogen sources on the growth and the lovastatin production of the mutant and the original strains were investigated comparatively. The spores of A. terreus CA99 were irradiated by 15, 20, 25, and 30 Gy of 80 MeV/u (12)C(6+) heavy-ion beams. Based on the lovastatin contents in the fermentation broth, a strain designated as A. terreus Z15-7 has been selected from the clone irradiated by the heavy-ion beam. When compared with the original strain, the content of lovastatin in the fermentation broth of A. terreus Z15-7 increased 4-fold. Moreover, A. terreus Z15-7 efficiently used the carbon and nitrogen sources for the growth and production of lovastatin when compared to the original strain. The maximum yield of lovastatin, 916.7 µg/ml, was obtained as A. terreus Z15-7 was submerged cultured in the chemically defined medium supplemented with 3% glycerol as a carbon source, 1% corn meal as an organic nitrogen source, and 0.2% sodium nitrate as an inorganic nitrogen source at 30 °C in the shake flask. The result shows that heavy-ion beam irradiation is an effective method for the mutation breeding of lovastatin production of A. terreus.

Identification and characterization of integron-associated antibiotic resistant Laribacter hongkongensis isolated from aquatic products in China.

Laribacter hongkongensis is a recently discovered bacterium associated with gastroenteritis. In this study, a total of 199 isolates of this species obtained from aquatic products (n=462) in Guangzhou City, China, were examined for their susceptibility to 19 antimicrobial agents and the presence of antimicrobial resistance integrons. The genetic relatedness of the isolates with integrons was also evaluated. A PCR-based method was used to screen integrons and found that 13 (6.5%) of the isolates harbored class 1 integrons. The antimicrobial resistance rates of integron-positive isolates were significantly higher than integron-negative ones for cefepime, ciprofloxacin, gentamicin, tetracycline, trimethoprim-sulfamethoxazole and rifampicin. Genetic sequence analysis revealed that these integrons contained various antimicrobial-resistance genes (dfrA1, dfrA14, dfrA17, dfrA32, aadA1, aadA2, aadA5, cmlA5, arr2, ereA and orfC) organized into different gene cassettes arrangements including a novel array of dfrA14-arr2-cmlA5. Pulsed-field gel electrophoresis (PFGE) yielded 13 different patterns among 13 integron-positive isolates, which could be grouped into four clusters. These indicate the dispersal of multi-resistant integrons among different molecular types. To the best of our knowledge, this is the first report showing distribution and characterization of class 1 integrons among L. hongkongensis isolates.