Advances in Dyslipidaemia Treatments: Focusing on ApoC3 and ANGPTL3 Inhibitors.

Apolipoprotein C3 (apoC3) and angiopoietin-like protein 3 (ANGPTL3) inhibit lipolysis by lipoprotein lipase and may influence the secretion and uptake of various lipoproteins. Genetic studies show that depletion of these proteins is associated with improved lipid profiles and reduced cardiovascular events so it was anticipated that drugs which mimic the effects of loss-of-function mutations would be useful lipid treatments. ANGPTL3 inhibitors were initially developed as a treatment for severe hypertriglyceridaemia including familial chylomicronaemia syndrome (FCS), which is usually not adequately controlled with currently available drugs. However, it was found ANGPTL3 inhibitors were also effective in reducing low-density lipoprotein cholesterol (LDL-C) and they were studied in patients with homozygous familial hypercholesterolaemia (FH). Evinacumab targets ANGPTL3 and reduced LDL-C by about 50% in patients with homozygous FH and it has been approved for that indication. The antisense oligonucleotide (ASO) vupanorsen targeting ANGPTL3 was less effective in reducing LDL-C in patients with moderate hypertriglyceridaemia and its development has been discontinued but the small interfering RNA (siRNA) ARO-ANG3 is being investigated in Phase 2 studies. ApoC3 can be inhibited by the ASO volanesorsen, which reduced triglycerides by >70% in patients with FCS and it was approved for FCS in Europe but not in the United States because of concerns about thrombocytopaenia. Olezarsen is an N-acetylgalactosamine-conjugated ASO targeting apoC3 which appears as effective as volanesorsen without the risk of thrombocytopaenia and is undergoing Phase 3 trials. ARO-APOC3 is an siRNA targeting apoC3 that is currently being investigated in Phase 3 studies.

Research trends in cardiovascular tissue engineering from 1992 to 2022: a bibliometric analysis.

Background: Cardiovascular tissue engineering (CTE) is a promising technique to treat incurable cardiovascular diseases, such as myocardial infarction and ischemic cardiomyopathy. Plenty of studies related to CTE have been published in the last 30 years. However, an analysis of the research status, trends, and potential directions in this field is still lacking. The present study applies a bibliometric analysis to reveal CTE research trends and potential directions. Methods: On 5 August 2022, research articles and review papers on CTE were searched from the Web of Science Core Collection with inclusion and exclusion criteria. Publication trends, research directions, and visual maps in this field were obtained using Excel (Microsoft 2009), VOSviewer, and Citespace software. Results: A total of 2,273 documents from 1992 to 2022 were included in the final analysis. Publications on CTE showed an upward trend from 1992 [number of publications (Np):1] to 2021 (Np:165). The United States (Np: 916, number of citations: 152,377, H-index: 124) contributed the most publications and citations in this field. Research on CTE has a wide distribution of disciplines, led by engineering (Np: 788, number of citations: 40,563, H-index: 105). "Functional maturation" [red cluster, average published year (APY): 2018.63, 30 times], "cell-derived cardiomyocytes" (red cluster, APY: 2018.43, 46 times), "composite scaffolds" (green cluster, APY: 2018.54, 41 times), and "maturation" (red cluster, APY: 2018.17, 84 times) are the main emerging keywords in this area. Conclusion: Research on CTE is a hot research topic. The United States is a dominant player in CTE research. Interdisciplinary collaboration has played a critical role in the progress of CTE. Studies on functional maturation and the development of novel biologically relevant materials and related applications will be the potential research directions in this field.

Current hotspot and study trend of innate immunity in COVID-19: a bibliometric analysis from 2020 to 2022.

Background: Since the coronavirus disease 2019 (COVID-19) has spread throughout the world, many studies on innate immunity in COVID-19 have been published, and great progress has been achieved, while bibliometric analysis on hotspots and research trends in this field remains lacking. Methods: On 17 November 2022, articles and reviews on innate immunity in COVID-19 were recruited from the Web of Science Core Collection (WoSCC) database after papers irrelevant to COVID-19 were further excluded. The number of annual publications and the average citations per paper were analyzed by Microsoft Excel. Bibliometric analysis and visualization of the most prolific contributors and hotspots in the field were performed by VOSviewer and CiteSpace software. Results: There were 1,280 publications that met the search strategy on innate immunity in COVID-19 and were published from 1 January 2020 to 31 October 2022. Nine hundred thirteen articles and reviews were included in the final analysis. The USA had the highest number of publications (Np) at 276 and number of citations without self-citations (Nc) at 7,085, as well as an H-index of 42, which contributed 30.23% of the total publications, followed by China (Np: 135, Nc: 4,798, and H-index: 23) with 14.79% contribution. Regarding Np for authors, Netea, Mihai G. (Np: 7) from the Netherlands was the most productive author, followed by Joosten, Leo A. B. (Np: 6) and Lu, Kuo-Cheng (Np: 6). The Udice French Research Universities had the most publications (Np: 31, Nc: 2,071, H-index: 13), with an average citation number (ACN) at 67. The journal Frontiers in Immunology possessed the most publications (Np: 89, Nc: 1,097, ACN: 12.52). "Evasion" (strength 1.76, 2021-2022), "neutralizing antibody" (strength 1.76, 2021-2022), "messenger RNA" (strength 1.76, 2021-2022), "mitochondrial DNA" (strength 1.51, 2021-2022), "respiratory infection" (strength 1.51, 2021-2022), and "toll-like receptors" (strength 1.51, 2021-2022) were the emerging keywords in this field. Conclusion: The study on innate immunity in COVID-19 is a hot topic. The USA was the most productive and influential country in this field, followed by China. The journal with the most publications was Frontiers in Immunology. "Messenger RNA," "mitochondrial DNA," and "toll-like receptors" are the current hotspots and potential targets in future research.

High-intensity and moderate-intensity interval training in heart failure with preserved ejection fraction: A meta-analysis of randomized controlled trials.

BACKGROUND: Exercise training significantly improves cardiorespiratory fitness (CRF) in heart failure with reduced ejection fraction (HFrEF) patients, but high-intensity interval training (HIIT) is not superior to moderate-intensity interval training (MIIT). Whether HIIT is more beneficial than MIIT in patients with heart failure with preserved ejection fraction (HFpEF) remains unclear. METHODS: On August 29, 2021, we conducted a comprehensive computerized literature search of the Medline, EMBASE, Web of Science, and Cochrane databases using the following keywords: "HF or diastolic HF or HFpEF or HF with normal ejection fraction and exercise training or aerobic exercise or isometric exercises or physical activity or cardiac rehabilitation." Only randomized controlled trials (RCTs) reporting comparisons between HIIT and MIIT in HFpEF were included in the final analysis to maintain consistency and obtain robust pooled estimates. Methodological quality was assessed based on the ratings of individual biases. To generate an overall test statistic, the data were analyzed using the random-effects model for a generic inverse variance. Outcome measures were reported as an odds ratio, and confidence intervals (CIs) were set at 95%. The study followed PRISMA guidelines. RESULTS: This meta-analysis included only RCTs comparing the efficacy of HIIT and MIIT in HFpEF patients. This study included 150 patients from 3 RCTs. In the current pooled data analysis, HIIT significantly improves diastolic function measured by E/A ratio (WMD, 0.13; 95% CI, 0.03-0.23, P = .009). However, no significant change was observed in the diastolic function measured by E/e' ratio (WMD, 0.39; 95% CI, -2.40 to 3.18, P = .78), and CRF evaluated by both VO2 (mL/kg per min; WMD, -0.86; 95%CI, -5.27 to 3.55, P = .70) and VE/CO2 slope (WMD, 0.15; 95% CI, -10.24 to 10.53, P = .98), and systolic function (EF-WMD, -2.39; 95% CI, -12.16% to 7.38%, P = .63) between HIIT and MIIT in patients with HFpEF. CONCLUSION: In HFpEF patients, HIIT may be superior to MIIT in improving diastolic function, measured by E/A, but not CRF and left ventricular systolic function.

Identification of potential immunomodulators from Pulsatilla decoction that act on therapeutic targets for ulcerative colitis based on pharmacological activity, absorbed ingredients, and in-silico molecular docking.

BACKGROUND: Pulsatilla decoction (Bai-Tou-Weng-Tang, BTWT) is a classic formula prescription of a traditional Chinese medicine that is used to treat ulcerative colitis (UC). However, its active components and underlying mechanism of action remain unclear. In the present study, we aimed to identify potential immunomodulators from BTWT that act at therapeutic targets for UC. METHODS: The protective effects of BTWT granules were examined in mice with colitis induced by dextran sulfate sodium. The absorbed components of BTWT were identified using LC-MS, and selected protein targets of these components in UC were investigated using molecular docking. RESULTS: Oral administration of BTWT granules significantly alleviated disease severity and colon shortening, and inhibited the inflammatory response in mice with chronic colitis. In these mice, 11 compounds from the BTWT granules were detected in the serum and/or colon. The molecular docking study demonstrated that compounds from Radix pulsatillae, such as anemoside A3, interacted with STAT3 and S1PR1; compounds from Rhizoma coptidis and/or Cortex phellodendri, such as palmatine, interacted with JAK3, PD-1, and PD-L1; and components of Cortex fraxini such as aesculin interacted with S1PR1, JAK3, STAT3 and PD-L1. Further in-vitro experiments showing that the compounds inhibited TNF-α and IL-6 production and STAT3 activation in RAW 264.7 cells suggested that these compounds have immunomodulatory activities. CONCLUSION: We revealed for the first time that 11 absorbed ingredients from BTWT were immunomodulators against therapeutic targets for UC. These findings suggest that the identified compounds are the active components of BTWT, and the identified protein targets underlie the mechanism of action of BTWT against UC.

Prevalence of Escherichia coli ST1193 Causing Intracranial Infection in Changsha, China.

ST1193 is an emerging new virulent and resistant clone among Escherichia coli with a tendency to spread rapidly across the globe. However, the prevalence of intracranial infection-causing E. coli ST1193 is rarely reported. This study aimed at determining the prevalence of E. coli ST1193 isolates, causing intracranial infections in Changsha, central China. A total of 28 E. coli isolates were collected from the cerebrospinal fluid of patients with intracranial infection over a four-year period. All isolates were differentiated using multilocus sequence typing (MLST), and phylogenetic grouping, and tested for antibiotic resistance. MLST analysis showed 11 sequence types (ST) among the 28 E. coli isolates. The most prevalent ST was B2-ST1193 (28.6%, 8/28), followed by B2-ST131 (21.4%, 6/28) and F-ST648 (10.7%, 3/28). Of the eight ST1193 isolates, three carried CTX-M-55, and one carried CTX-M-27. All eight ST1193 isolates were resistant to Ciprofloxacin, showing gyrA1AB/parC4A mutations. Two ST1193 isolates carried the aac(6')-Ib-cr gene. All ST1193 isolates were recovered from infants with meningitis, with a fatal outcome for one three-month-old infant. ST1193 has emerged as the predominant type of E. coli strain causing intracranial infections in Changsha, China. This study highlights the importance of implementing appropriate surveillance measures to prevent the spread of this emerging public health threat.

Emerging trends in sacubitril/valsartan research: A bibliometric analysis of the years 1995-2021.

BACKGROUND: Sacubitril/valsartan has been approved for the treatment of heart failure (HF) patients with reduced ejection fraction; since then, it gradually became a new star drug in the therapy of HF. Nevertheless, the effectiveness of sacubitril/valsartan remains under investigation. Thus far, only a few bibliometric studies have systematically analyzed the application of sacubitril/valsartan. METHODS: Publications on sacubitril/valsartan were retrieved from the Web of Science Core Collection on April 29, 2021. Data were analyzed using Microsoft Excel 2019 (Redmond, WA), VOS viewer (Redmond, WA), and Cite Space V (Drexel University, Philadelphia, PA). RESULTS: A total of 1309 publications on sacubitril/valsartan published from 1995 to 2021 were retrieved. The number of publications regarding sacubitril/valsartan increased sharply in the last 6 years (2015-2021), and American scholars authored >40% of those publications. Most were published in the European Journal of Heart Failure, the United States was the bellwether with a solid academic reputation in this area. Solomon published the highest number of related articles and was the most frequently cited author. "Heart failure" was the leading research hotspot. The keywords, "inflammation," "fibrosis," and "oxidative stress" appeared most recently as research fronts. CONCLUSIONS: Research attention should be focused on clinical trial outcomes. Considering its effectiveness in HF, the mechanisms and further applications of sacubitril/valsartan may become research hotspots in the future and should be closely examined.

Community Fecal Carriage and Molecular Epidemiology of Extended-Spectrum ß-Lactamase- and Carbapenemase-Producing Escherichia coli from Healthy Children in the Central South China.

Background: Fecal carriage of extended-spectrum ß-lactamase-producing Escherichia coli (ESBL-EC) and carbapenemase-producing E. coli (CP-EC) is well reported among hospitalized adults and children. However, there are few studies on the carriage prevalence and ESBL-EC and CP-EC genotypes among healthy children in China. Patients and Methods: Stool samples were collected from 330 students in 2021 from three randomly selected primary schools in Changsha, China. ESBL-EC and CP-EC were screened using CHROMagarTM chromogenic plates. ESBL and carbapenemase production was confirmed using the double-disc synergy test and a modified carbapenem inactivation method, respectively. Antimicrobial susceptibility was tested using the broth microdilution method. Resistance determinants, virulence factors, and phylogenetic groups were determined by PCR and sequencing. Multi-locus sequence typing (MLST) was performed (seven housekeeping genes were amplified and sequenced) on the phylogenic group B2 E. coli to detect high-risk clonal strains such as ST131 E. coli. Then, ST131 E. coli were characterized based on ST131 clades, O-type, and fimH alleles. Results: In total, 118 (35.8%) ESBL-EC and 3 (0.9%) CP-EC were isolated. bla CTX-M was the most common genotype (27.1%), identified in all ESBL-EC, except one, which carried bla SHV-12. One isolate with mcr-1 was found amongst ESBL-EC, whereas all three CP-EC carried bla NDM-1. The predominant sequence type (ST) clones in group B2 were ST131 and ST1193. The prevalence of ST131 E. coli was 9.9%, displaying serotypes O16 and O25b, fimH alleles 30, 41, and 89, and ST131 clades A and C1-M27. Conclusion: In this study, high carriage rate of ESBL-EC was found among healthy children, and the dominant ESBL was CTX-M-14. In addition, high-risk clones (ST131 and ST1193) were also detected. This emphasizes the importance of monitoring ESBL-EC in community settings.

Lympho-vascular invasion impacts the prognosis in breast-conserving surgery: a systematic review and meta-analysis.

BACKGROUND: It is estimated that breast cancer (BC) incidence, especially that of early-stage breast cancer cases continues to rise due to increased universal screening. Breast-conserving surgery (BCS) is the main intervention for early-stage BC. Lympho-vascular invasion (LVI) is reported to influence breast cancer prognosis but its prognostic value in breast-conserving treatment is controversial. METHODS: A search was conducted on the Cochrane library, PubMed, Web of Science, and EMBASE from inception to December 1st, 2021, without language restrictions, to identify studies that explored the prognosis of lympho-vascular invasion in breast-conserving surgery. Reviews of each study were conducted, and data extracted. The meta-analysis was performed with StataSE 16. Study quality assessment was evaluated using the Newcastle-Ottawa Scale. RESULTS: Overall, 15 studies with 21,704 patients deemed eligible for this study. Event-free survival (EFS), disease-free survival (DFS), overall survival (OS), distant metastases (DM), loco-regional recurrence (LRR), local recurrence (LR), breast recurrence (BR), disease specific survival (DSS), and breast cancer specific survival (BCSS), were extracted from each study. We found that LVI leads to poor OS (HR = 1.46, 95% CI: 1.17-1.83), DM (HR = 2.08, 95% CI: 1.66-2.60) and LR (HR = 2.00, 95% CI: 1.54-2.61). CONCLUSIONS: We confirmed that early-stage BC patients with LVI-positive have poorer OS, DFS, LRR, BCSS, DM and LR following receiving BCS than those LVI-negative patients. Mastectomy, in combination with radical systemic therapies could be considered, especially in those requiring second surgery. How to change the impact of LVI on the local recurrence rate and long-term survival in patients who undergo BCS may be a valuable research direction in the future.

A bibliometric analysis of studies on the gut microbiota in cardiovascular disease from 2004 to 2022.

Background: Increasing evidence indicates that the gut microbiota (GM) is linked to cardiovascular disease (CVD). Many studies on the GM in CVD have been published in the last decade. However, bibliometric analysis in this field is still lacking. Methods: On 30 September 2022, a search of the Web of Science™ (WoS; Clarivate™, Philadelphia, PA, USA) yielded 1,500 articles and reviews on the GM and CVD. Microsoft Excel and CiteSpace and VOSviewer software were used to analyze publication trends and research hotspots in this field. Results: Our search generated 1,708 publications on the GM in CVD published between 2004 and 2022, and 1,500 articles and review papers were included in the final analysis. The number of publications relating to the GM in CVD increased from 1 in 2004 to 350 in 2021. China (485 publications, 9,728 non-self-citations, and an H-index of 47) and the USA (418 publications, 24,918 non-self-citations, and an H-index of 82) contributed 32.31%, and 27.85%, respectively, of the total number of publications. Examination of the number of publications (Np) and number of citations, excluding self-citations (Nc), of individual authors showed that Y. L. Tian (Np: 18, Nc: 262, and H-index: 12), from China, is the most productive author, followed by R. Knight (Np: 16, Nc: 3,036, and H-index: 15) and M. Nieuwdorp (Np: 16, Nc: 503, and H-index: 9). The Chinese Academy of Medical Sciences and Peking Union Medical College accounted for the largest number of publications (Np: 62, Nc: 3,727, and H-index: 13, average citation number (ACN): 60.11). The journal Nutrients had the most publications (Np: 73, Nc: 2,036, and ACN: 27.89). The emerging keywords in this field were "monooxygenase 3" (strength 3.24, 2020-2022), "short-chain fatty acid" (strength 4.63, 2021-2022), "fatty liver disease" (strength 3.18, 2021-2022), "metabolic disease" (strength 3.04, 2021-2022), "Mediterranean diet" (strength 2.95, 2021-2022), "prevention" (strength 2.77, 2021-2022), and "intestinal barrier" (strength 2.8, 2021-2022). Conclusion: Publications on the GM in CVD rapidly increased in the last decade. The USA was the most influential country in publications in this field, followed by China. The journal with the most publications was Nutrients. Monooxygenase-3, short-chain fatty acids, fatty liver disease, metabolic disease, the Mediterranean diet, intestinal barrier, and prevention are the current hotspots or potential hotspots for future study.

Efficacy of Enhanced Cytokine-Induced Killer Cells as an Adjuvant Immunotherapy for Renal Cell Carcinoma: Preclinical and Clinical Studies.

Cytokine-induced killer (CIK) cells have been proved to be an effective method of tumor immunotherapy in numerous preclinical and clinical studies. In our previous study, a new method was developed to prime and propagate CIK cells by the combination of IL-2 and IL-15, and this kind of CIK cells had enhanced antitumor effect on lung cancer. For renal cell carcinoma (RCC), immunotherapy plays an important role because of the poor efficacy of radiotherapy and chemotherapy. In this study, we further evaluated the antitumor effects of these enhanced CIK cells against RCC. Enhanced CIK cells were generated by IL-2 combined with IL-15 and identified by flow cytometry. HEK-293 and ACHN cell lines were used to verify the efficiency of CIK cells in vitro, and then the ACHN tumor xenograft model was also employed for in vivo study. In addition, the secreted cytokines including IFN-γ, granzyme B, TNF-α, and perforin, as well as the local microstructure were also studied. Subsequently, 20 patients with RCC were enrolled into our study, and 11 patients were randomly divided into the autologous CIK treatment group for clinical research. The results showed that enhanced CIK cells exert better antitumor effects in RCC in vitro (p < 0.01 in HEK-293 and p < 0.05 in ACHN）and in vivo (p < 0.05). Patients benefit overall survival from enhanced CIK therapy in our clinical study. Our present preclinical and clinical studies for the first time elucidated that these enhanced CIK cells would be used as an effective adjuvant therapy in the treatment of RCC.

Reexamining the Solar Axion Explanation for the XENON1T Excess.

The XENON1T collaboration has observed an excess in electronic recoil events below 5 keV over the known background, which could originate from beyond-the-standard-model physics. The solar axion is a well-motivated model that has been proposed to explain the excess, though it has tension with astrophysical observations. The axions traveling from the Sun can be absorbed by the electrons in the xenon atoms via the axion-electron coupling. Meanwhile, they can also scatter with the atoms through the inverse Primakoff process via the axion-photon coupling, which emits a photon and mimics the electronic recoil signals. We found that the latter process cannot be neglected. After including the keV photon produced via the inverse Primakoff process in the detection, the tension with the astrophysical constraints can be significantly reduced. We also explore scenarios involving additional new physics to further alleviate the tension with the astrophysical bounds.

Testing the Sensitivity of the Galactic Center Excess to the Point Source Mask.

The Fermi Large Area Telescope (Fermi-LAT) Collaboration has an updated point source catalog, referred to as 4FGL. We perform the first template fit using a mask based on this new catalog and find that the excess in gamma rays detected at the Galactic Center in Fermi-LAT data persists. On the other hand, we find that a search for point sources is highly sensitive to the use of the 4FGL catalog: no sizable excess of bright pixels is apparent in the inner Galaxy when we mask out 4FGL point sources. Combining these observations restricts the ability of point sources to contribute to the Galactic Center excess. After identifying which bright sources have no known counterpart, we place strong constraints on any point source luminosity function capable of explaining the smooth emission identified in the template fit.

Constraining Dissipative Dark Matter Self-Interactions.

We study the gravothermal evolution of dark matter halos in the presence of dissipative dark matter self-interactions. Dissipative interactions are present in many particle-physics realizations of the dark-sector paradigm and can significantly accelerate the gravothermal collapse of halos compared to purely elastic dark matter self-interactions. This is the case even when the dissipative interaction timescale is longer than the free-fall time of the halo. Using a semianalytical fluid model calibrated with isolated and cosmological N-body simulations, we calculate the evolution of the halo properties-including its density profile and velocity dispersion profile-as well as the core-collapse time as a function of the particle model parameters that describe the interactions. A key property is that the inner density profile at late times becomes cuspy again. Using 18 dwarf galaxies that exhibit a corelike dark matter density profile, we derive constraints on the strength of the dissipative interactions and the energy loss per collision.

Escherichia coli O25b-ST131 and O16-ST131 causing urinary tract infection in women in Changsha, China: molecular epidemiology and clinical characteristics.

OBJECTIVE: This study aimed to investigate the prevalence of Escherichia coli ST131 and molecularly characterize the O25b-ST131 and O16-ST131 subgroups among urinary tract infection (UTI) E. coli isolates from women in central China. We also assessed the clinical characteristics and outcomes of infections caused by E. coli ST131. METHODS: Between January 2014 and December 2015, a total of 216 consecutive, non-repetitive E. coli isolates were recovered from UTI urine samples from women in Changsha, China. All isolates were analyzed for phylogenetic groups, antimicrobial resistance and virulence genotypes. ST131 clonal groups were identified using PCR and characterized using O serotyping, CTX-M genotypes, fimH, gyrA, and parC alleles, fluoroquinolone resistance genes and pulsed-field gel electrophoresis (PFGE). Clinical data were obtained from medical records. RESULTS: Overall, 41 (19.0%) of 216 E. coli isolates were identified to contain ST131 strains, among which 27 were O25b-ST131 strains and 14 were O16-ST131 strains. The clinical characteristics and outcomes of the ST131 group did not differ significantly from those of the non-ST131 group, except for the presence of urinary stones (43.9% vs 27.4%, P=0.039). Ciprofloxacin resistance was found to be significantly higher in O25b-ST131 isolates than O16-ST131 isolates (96.3% vs 14.3%, P<0.001). The majority of O25b-ST131 isolates belonged to fimH30 (92.6%), followed by fimH41 (3.7%) and fimH27 (3.7%). O25b-H30 and O25b-H41 isolates were resistant to ciprofloxacin, and possessed gyrA1AB/parC1aAB combination. All of the O16-S131 isolates were found to belong to fimH41, and of which, two of the ciprofloxacin-resistant strains harbored gyrA1AB/parC3A combination. Three PFGE clusters, consisting of 38 (92.7%) isolates, with more than 70% similarity were identified. CONCLUSION: The O25b and O16 sub-lineages have emerged as an important group of E. coli ST131 in UTI isolates from women in China. UTI patients with a history of urinary stones may need to be particularly vigilant against ST131 infection.

Epidemiology and molecular characterization of mcr-1 in Escherichia coli recovered from patients with bloodstream infections in Changsha, central China.

OBJECTIVES: The main aim of this study was to investigate the prevalence and molecular characteristics of the mcr-1 gene in Escherichia coli isolates obtained from all patients with bloodstream infections over a year in a Chinese teaching hospital. We also assessed the susceptibility profiles of the mcr-1-positive strains and prognostic impact of this gene on the patients. METHODS: A total of 144 consecutive, non-repetitive E. coli isolates causing bloodstream infections were collected at a teaching hospital in Changsha, China from January to December 2016. The presence of the mcr-1 gene was assessed by PCR. All mcr-1-positive E coli isolates were characterized by antimicrobial susceptibility testing, multilocus sequence typing (MLST), a conjugation experiment, and plasmid replicon typing. Clinical data were obtained from medical records. RESULTS: The mcr-1 gene was detected in three (2.1%) of the 144 E. coli isolates. The three mcr-1-positive E. coli isolates were resistant to colistin. All three isolates showed a lower resistance to other classes of antibacterials, with all three being susceptible to carbapenems. The MLST results indicated that the three E. coli isolates were assigned to three different sequence types: ST457, ST101, and ST1413, respectively. The conjugation experiment showed that the mcr-1 gene was successfully transferred to the recipient (E. coli EC600) from two isolates, one of which possessed IncI1 replicons and the other of which carried IncHI2 and IncN replicons. The patients with bloodstream infections caused by mcr-1-positive isolates had severe underlying diseases and were cured after antibacterial treatment. CONCLUSION: The prevalence of the mcr-1 gene in patients with E. coli bloodstream infection was 2.1% in Changsha, China. The mcr-1-positive E. coli isolates had varied susceptibility profiles, although all three were susceptible to carbapenems. This therapeutic window is crucial given the risk of rapid deterioration in high-incidence areas worldwide.

[Huanshao Capsules combined with levocarnitine for the treatment of asthenospermia, oligospermia and teratozoospermia].

OBJECTIVE: To observe the clinical effects of Huanshao Capsules (HSC) combined with levocarnitine (LC) on asthenospermia, oligospermia, teratozoospermia, and the semen parameters of the patients. METHODS: This randomized controlled clinical study included 186 infertility patients with spleen and kidney asthenia. We randomly divided them into three groups of equal number and treated them orally with HSC at the dose of 3 capsules tid, LC at 10 ml tid, and HSC+LC, respectively, all for 12 weeks. At 4, 8, and 12 weeks after treatment, we obtained the semen parameters from the patients and compared them among the three groups. RESULTS: Totally, 180 of the patients completed the study, 61 in the HSC, 59 in the LC and 60 in the HSC+LC group. After 12 weeks of medication, the patients of the HSC+LC group showed an increase of 42.77% in the semen volume, 142.37% in sperm concentration, 28.61% in sperm motility, and 24.39% in the percentage of grade a+b sperm and a decrease of 6.27% in the percentage of morphologically abnormal sperm as compared with the baseline (P <0.05). The patients treated with HSC+LC showed significantly more improvement in all the above parameters than those treated with LC alone (P <0.05) as well as in sperm motility and the percentage of progressively motile sperm than those treated with HSC alone (P <0.05). The HSC group exhibited remarkable improvement in the above parameters after treatment as compared with the baseline (P <0.05) and higher semen volume and sperm concentration than the LC group (P <0.05). CONCLUSIONS: Huanshao Capsulescombined with levocarnitinedeserves a wide clinical application as a safe and efficacious therapy forasthenospermia, oligospermia,and teratozoospermia.

[Emergence of methicillin-resistant Staphylococcus aureus SCCmec type IV/V epidemic clones in a large teaching hospital in China].

OBJECTIVE: To investigate the staphylococcal cassette chromosome mec (SCCmec) genotype and molecular epidemiological characteristics of healthcare-associated methicillin-resistant Staphylococcus aureus (HA-MRSA) in a large teaching hospital in China. METHDOS: From January 2012 to December 2012, a total of 71 nonduplicate HA-MRSA were collected in a teaching hospital in Changsha, China. SCCmec types were determined by multiplex PCR, and Panton-Valentine leukocidin (PVL) gene was detected by PCR. The homology among the tested isolates was determined using pulsed-field gel electrophoresis (PFGE). RESULTS: Of the 71 HA-MRSA isolates, 49 (69.0%) carried SCCmec III, 10 (14.1%) carried SCCmec IV, 3 (4.2%) carried SCCmec V and 3 (4.2%) carried SCCmec II; the remaining 6 isolates were not typeable by PCR. Compared with patients having SCCmec I/II/III MRSA infections, those with SCCmec IV/V MRSA infections had a significantly younger age and a similar duration of hospital stay before the first MRSA-positive culture and total hospital stay. PVL genes were strongly associated with SCCmec type IV/V MRSA infections. HA-SCCmec IV/V MRSA strains showed a greater susceptibility to rifampicin, gentamicin, levofloxacin, ciprofloxacin, and tetracycline than HA-SCCmec I/II/III MRSA strains. The 13 HA-SCCmec IV/V MRSA isolates formed one large group at the 55% similarity level. Three PFGE clusters with a similarity index of 85% or more were identified, and unique PFGE profiles were observed in 4 isolates. CONCLUSION: This is the first report of HA-MRSA isolates carrying SCCmec V in Chinese hospitals. SCCmec types IV and V MRSA clones have emerged in Chinese hospitals, which urges more rigorous surveillance of their spread in healthcare facilities in China.

Preparation and preliminary application of monoclonal antibodies to the receptor binding region of Clostridium difficile toxin B.

A previous nationwide Chinese epidemiological study revealed through isolation of A­B+ Clostridium difficile strains, which produce toxin B (TcdB), but not toxin A TcdA, that the strains are widespread and more frequent in east Asian countries,. The development of a process capable of detecting TcdB is required in microbiological laboratories in order to facilitate the control of the A­B+ C. difficile strains, however, no diagnostic reagents have been developed to date. The aim of the present study was to prepare monoclonal antibodies (mAbs) targeting the receptor binding region of TcdB (CDB3), and to establish a double­antibody sandwich enzyme-linked immunosorbent assay (ds­ELISA), which can be used for the diagnosis of C. difficile infection. The recombinant protein, glutathione S transferase (GST)­CDB3 was expressed and purified using an Escherichia coli system. BALB/c mice were immunized with GST­CDB3 recombinant protein. A hybridoma technique was used for the production of anti­CDB3 mAb. The hybridoma clones were then screened using indirect ELISA, and anti­CDB3 mAb was produced in the ascites of the BALB/c mice. Isotyping of anti­CDB3 mAb was performed using an SBA Clonotyping system/horseradish peroxidase (HRP) ELISA kit. Protein G affinity chromatography was used for purification of anti­CDB3 mAbs, and the titer and specificity of the anti­CDB3 mAbs were assessed using indirect ELISA and western blot analysis, respectively. The ds­ELISA was established using HRP­labeled anti­CDB3 mAbd, which were used to detect TcdB clinically in diarrhea stools. A total of five stable hybridoma cell clones (1E7B, 1F8D3, 2F8A6, 3B6F1 and 4A4G2) producing anti­CDB3 mAb were established. The results of the present study indicated that the immunoglobulin (Ig)G isotype was predominant, as 1E7B2 IgG1 (λ), 2F8A6 IgG2a (κ) and 4A4G2 IgG1 (κ). In addition, the highest titer of anti­CDB3 mAb (2F8A6 and 4A4G2) was 1:51,200. Western blotting revealed that the 2F8A6 and 4A4G2 mAbs recognized the CDB3 protein specifically. Following anti­CDB3 mAb (4A4G2) HRP­labeling, the optimal working concentration was confirmed to be 1:400, and the concentration of coated antibody (2F8A6) was 20 µg/ml. The sensitivity of the ds­ELISA was 73.33% for the A+B+ toxigenic C. difficile strains, and 86.67% for the A­B+ toxigenic C. difficile strains, with a specificity of 100% for all. In conclusion, the present study successfully developed novel mAbs specific to CDB3, and developed a ds-ELISA kit with high specificity and sensitivity for the rapid detection of TcdB. This offers a useful tool for the diagnostic assessment of TcdB.

Emergence and spread of O16-ST131 and O25b-ST131 clones among faecal CTX-M-producing Escherichia coli in healthy individuals in Hunan Province, China.

OBJECTIVES: The objectives of this study were to determine CTX-M-producing Escherichia coli ST131 strain prevalence in stool specimens from healthy subjects in central China and to molecularly characterize clonal groups. METHODS: From November 2013 to January 2014, stool specimens from healthy individuals in Hunan Province were screened for ESBL-producing E. coli using chromogenic medium and CTX-M genotypes and phylogenetic groups were determined. ST131 clonal groups were detected by PCR and characterized for antibiotic resistance, fimH, gyrA and parC alleles, plasmid-mediated quinolone resistance determinants, virulence genotypes and PFGE patterns. RESULTS: Among 563 subjects, 287 (51.0%) exhibited the presence of faecal ESBL-producing E. coli, all of which produced CTX-M enzymes. The most common CTX-M genotypes were CTX-M-14 (48.4%), CTX-M-15 (27.5%) and CTX-M-27 (15.0%). Of the 287 CTX-M-producing isolates, 32 (11.1%) belonged to the ST131 clone. O16-ST131 isolates were dominant (75%) and contained the fimH41 allele. The remaining eight (25%) ST131 isolates were of the O25b subgroup and contained fimH30 or fimH41. Ciprofloxacin resistance was found in 100% of the O25b-ST131 isolates, whereas only 8% of the O16-ST131 isolates were resistant. All of the O25b-ST131 isolates except one showed gyrA1AB and parC1aAB mutations; most of the O16-ST131 isolates had gyrA1A and parC1b mutations. The virulence genotypes of O16-ST131 resembled those of the O25b-ST131 isolates. The 32 ST131 isolates formed one large group at the 64% similarity level. They comprised 15 PFGE groups (defined at ≥85% similarity). CONCLUSIONS: O16-ST131 isolates have emerged as the predominant type of ST131 isolate in faecal CTX-M-producing E. coli in healthy individuals in China.