[Establishment and application of a multilocus sequence typing assay for Corynebacterium striatum].

Objective: To establish a multilocus sequence typing (MLST) assay for Corynebacterium (C.) striatum, explore the population structure and evolution relationship of clinical isolates of C. striatum. Methods: Seven housekeeping genes (gyrA, gyrB, hsp65, sodA, secA1, rpoB, 16S rRNA) were amplified with PCR by using self-designed specific primers and sequenced. Then, the sequences were assembled with software SeqMan. The gene diversity and gene recombination characteristics were evaluated by using software DnaSP 5.10.01 and Splits tree 4.14.2. The phylogenetic tree and the minimum spanning tree were constructed based on the sequence types (ST) characteristics by using software MEGA 7.0.14 and BioNumerics, respectively. In addition, the genetic evolutionary relationship among STs were analyzed by using software eBURST 3.0. Results: The expected amplification products of seven sites selected in all the test strains were obtained. Splits tree showed that the clustering of all C. striatum strains was consistent, suggesting that gene recombination is the potential driving force for the evolution of C. striatum. All of the 344 C.striatum strains were divided into 72 STs by MLST and 85.7% of the strains formed clonal complexes. CC19 was the predominant clonal complex, whereas ST16 in the clonal complex was detected in the most strains. ST had a certain geographic clustering and a certain correlation with the isolation time. Conclusions: C. striatum showed high genetic diversity in China and CC19 was the predominant clonal complex. The MLST assay established in this study can be used for the typing of C. striatum, but further improvement is needed.

Effect miR-214-3p on proliferation and apoptosis of breast cancer cells by targeting survivin protein.

OBJECTIVE: Studies showed that microRNAs (miRs) play an important role in the development of breast cancer. It has been shown that there were significant differences between the expression levels of serum miR-214-3p in breast cancer patients and healthy controls. Since survivin is involved in cell cycle and apoptosis, this study aims to investigate the effect of miR-214-3p on the proliferation and apoptosis of breast cancer cells. MATERIALS AND METHODS: Dual-Luciferase reporter system was used to validate the cell cycle-related target gene survivin. miRanda and TargetScan were used to predict miR-214-3p target genes. Lipofectamine 2000 was used to transfect the miR-214-3p mimics, miR-NC into the MCF-7 cells. The quantitative Real Time-PCR (qRT-PCR) was used to detect the expression levels of miR-214-3p and survivin. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to examine the cell proliferation of breast cancer cells. The flow cytometry assay was used to evaluate the apoptosis of breast cancer cells. RESULTS: Dual-Luciferase reporter assay showed that cells co-transfected with wild-type vector and miR-214-3p mimics had significant lower ratios of hRluc/Luc fluorescence compared to that of the control group (p<0.05). The expression level of miR-214-3p was increased along with the increase of time after transfection, whereas the expression level of survivin mRNA was decreased along with the increase of time post transfection. This result suggests that miR-214-3p regulates the mRNA expression of survivin. Transfection of miR-214-3p inhibitor increased the proliferation of MCF-7 cells and transfection of miR-214-3p mimics decreased the proliferation of MCF-7 cells compared to control group (p<0.05). CONCLUSIONS: Survivin gene is a downstream target of miR-214-3p in breast cancer cells. The expression of miR-214-3p and survivin is correlated with the proliferation and apoptosis of breast cancer cells.

Perioperative outcomes and hospital reimbursement by type of radical prostatectomy: results from a privately insured patient population.

BACKGROUND: With the increasing use of robotic surgery in the United States, the comparative effectiveness and differences in reimbursement of minimally invasive radical prostatectomy (MIRP) and open prostatectomy (ORP) in privately insured patients are unknown. Therefore, we sought to assess the differences in perioperative outcomes and hospital reimbursement in a privately insured patient population who were surgically treated for prostate cancer. METHODS: Using a large private insurance database, we identified 17,610 prostate cancer patients who underwent either MIRP or ORP from 2003 to 2010. The primary outcomes were length of stay (LOS), perioperative complications, 90-day readmissions rates and hospital reimbursement. Multivariable regression analyses were used to evaluate for differences in primary outcomes across surgical approaches. RESULTS: Overall, 8981 (51.0%) and 8629 (49.0%) surgically treated prostate cancer patients underwent MIRP and ORP, respectively. The proportion of patients undergoing MIRP markedly rose from 11.9% in 2003 to 72.5% in 2010 (P<0.001 for trend). Relative to ORP, MIRP was associated with a shorter median LOS (1.0 day vs 3.0 days; P<0.001) and lower adjusted odds ratio of perioperative complications (OR: 0.82; P<0.001). However, the 90-day readmission rates of MIRP and ORP were similar (OR: 0.99; P=0.76). MIRP provided higher adjusted mean hospital reimbursement compared with ORP (US $19,292 vs. US $17,347; P<0.001). CONCLUSIONS: Among privately insured patients diagnosed with prostate cancer, robotic surgery rapidly disseminated with over 70% of patients undergoing MIRP by 2009-2010. Although MIRP was associated with shorter LOS and modestly better perioperative outcomes, hospitals received higher reimbursement for MIRP compared with ORP.

Specialty bias in treatment recommendations and quality of life among radiation oncologists and urologists for localized prostate cancer.

BACKGROUND: Given the importance of physician attitudes about different treatments and the quality of life (QOL) in prostate cancer, we performed a national survey of specialists to assess treatment recommendations and perceptions of treatment-related survival and QOL. METHODS: We mailed a self-administered survey instrument to a random sample of 1366 specialists in the U.S. Respondents were asked for treatment recommendations and survival that varied by PSA levels and Gleason scores and estimate QOL outcomes. Pearson's chi-square and multivariable regression models were used to test for differences in each outcome. RESULTS: Response rates were similar for radiation oncologists (52.6%) and urologists (52.3%; P=0.92). Across all risk strata, urologists were more likely to recommend surgery than were radiation oncologists, for conditions ranging from PSA>20 and Gleason score 8-10 (35.2 vs. 0.2%; P<0.001) to PSA 4-10 and Gleason score 7 (87.5 vs. 20.9%; P<0.001). Radiation oncologists were also more likely to recommend radiation therapy relative to urologists (all P<0.001). From low- to high-risk prostate cancer, radiation oncologists and urologists perceived their treatment as being better for improving survival (all P<0.001). Each specialty also viewed their treatment as having less urinary incontinence (all P<0.001). CONCLUSIONS: Radiation oncologists and urologists both prefer the treatment modalities they offer, perceive them to be more effective and to lead to a better QOL. Patients may be receiving biased information, and a truly informed consent process with shared decision-making may be possible only if they are evaluated by both specialties before deciding upon a treatment course.

Potential bias in the bank: what distinguishes refusers, nonresponders and participants in a clinic-based biobank?

BACKGROUND: Biobanks are an important resource for genetic and epidemiologic research, but bias may be introduced if those who accept the recruitment invitation differ systematically from those who do not in terms of attributes important to health-related investigations. To understand potential bias in a clinic-based biobank of biological samples, including genetic data linked to electronic health record information, we compared patient characteristics and self-reported information among participants, nonresponders and refusers. We also compared reasons for nonparticipation between refusers and nonresponders to elucidate potential pathways to reduce nonparticipation and any uncovered bias. METHODS: We mailed recruitment packets to 1,600 adult patients with upcoming appointments at Mayo Clinic (Rochester, Minn., USA) and recorded their participation status. Administrative data were used to compare characteristics across groups. We used phone interviews with 26 nonresponders and 26 refusers to collect self-reported information, including reasons for nonparticipation. Participants were asked to complete a mailed questionnaire. RESULTS: We achieved 26.2% participation (n=419) with 12.1% refusing (n=193) and 61.8% nonresponse (n=988). In multivariate analyses, sex, age, region of residence, and race/ethnicity were significantly associated with participation. The groups differed in information-seeking behaviors and research experience. Refusers more often cited privacy concerns, while nonresponders more often identified time constraints as the reason for nonparticipation. CONCLUSION: For genomic medicine to advance, large, representative biobanks are required. Significant associations between patient characteristics and nonresponse, as well as systematic differences between refusers and nonresponders, could introduce bias. Oversampling or recruitment changes, including heightened attention to privacy protection and participation burden, may be necessary to increase participation among less-represented groups.

Therapeutic effects of intravenous infusion of hyperoxygenated solution on acute haemorrhagic shock in rabbits.

Tissue anoxia is the main mechanism of the shock reaction. Here, the effect of hyperoxygenated solution (HOS) on acute haemorrhagic shock was studied in rabbits. At 60 min after shock, rabbits were infused intravenously with hyperoxygenated solution at 10 (HOS1 group) or 20 ml/kg (HOS2 group) or with Ringer's solution at 10 ml/kg (RS group). Compared with values before shock, values after shock were lower for mean arterial pressure (MAP), more negative for base excess (BE) and higher for blood lactate (BL) and blood viscosity. After infusion, MAP declined more slowly in the HOS1 and HOS2 groups than in the RS group. At 30 and 60 min after infusion, arterial partial pressure of oxygen (PaO(2)) and oxygen saturation (SaO(2)) were higher and BE was less negative in the HOS1 and HOS2 groups than in the RS group, BL was lower in the HOS1 and HOS2 groups than in the RS group, and PaO(2) and SaO(2) were higher in the HOS2 group than in the HOS1 group. It was concluded that HOS infusion can rectify changes in vital signs more effectively than Ringer's solution after acute haemorrhagic shock in rabbits.

Exon sequencing and association analysis of polymorphisms in TCF7L2 with type 2 diabetes in a Chinese population.

AIMS/HYPOTHESIS: Recently, variants in the transcription factor 7-like 2 (TCF7L2) gene have been found to be consistently associated with type 2 diabetes in different populations. In this study, we hypothesized that TCF7L2 also contributed to genetic susceptibility for type 2 diabetes in a Chinese population. METHODS: We looked for new variants by direct sequencing of all exons and intron-exon junctions of TCF7L2 in 100 Chinese type 2 diabetic patients, and then we genotyped five single nucleotide polymorphisms (SNPs) by Snapshot technology in 1,000 Chinese individuals. RESULTS: By sequencing, we identified six SNPs (c.1,637C>A; c.1,674C>G; c.1,709G>A; c.1,846C>G; c.1,888C>T; and c.1,876T>G), and three of them led to non-synonymous polymorphisms (c.1,637C>A, His-->Gln or Pro-->Thr; c.1,674C>G, Pro-->Arg; and c.1,709G>A, Ala-->Thr). All of them are rare except c.1,637C>A, which had a frequency of 0.23 for the minor A allele in 98 sequenced individuals. In a case-control study, one of the newly discovered SNPs (c.1,637C>A), together with four reported ones (rs7903146, rs12255372, rs290487 and rs3814573) were genotyped. Comparison between allele and genotype frequencies of these SNPs in patients and controls showed marginal association for rs7903146 and rs290487 with type 2 diabetes (p = 0.063, OR 1.982, 95% CI 1.128-3.485; p = 0.071, OR 1.237, 95% CI 0.983-1.557, respectively). No association was found for rs12255372, rs3814573, c.1,637C>A and type 2 diabetes (p = 0.278-1.000). CONCLUSIONS/INTERPRETATION: With the current sample size, we did not find any mutation in the coding sequence of TCF7L2 that confers a genetic risk for type 2 diabetes in a Chinese population, and did not replicate some of the major positive results obtained in other populations.

Mechanical strain induces osteogenic differentiation: Cbfa1 and Ets-1 expression in stretched rat mesenchymal stem cells.

Distraction osteogenesis is an active process of bone regeneration under controlled mechanical stimulation. Osteogenic differentiation of mesenchymal stem cells (MSCs) is essential for bone formation during this process. Cbfa1 and Ets-1 (core binding factor alpha 1 and v-ets erythroblastosis virus E26 oncogene homolog 1) are transcription factors that play important roles in the differentiation of MSCs to osteoblasts. In order to mimic a single activation of a clinical distraction device, a short period of cyclic mechanical strain (40 min and 2,000 microstrains) was applied to rat MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity were examined. The mRNA expression of Cbfa1 and Ets-1, as well as ALP, a specific osteoblast marker, was detected using real-time quantitative reverse transcription polymerase chain reaction. The results showed that mechanical strain can promote MSC proliferation, increase ALP activity and up-regulate the expression of Cbfa1 and Ets-1. A significant increase in Ets-1 expression was detected immediately after mechanical stimulation, but Cbfa1 expression was elevated later. The temporal expression pattern of ALP coincided perfectly with that of Cbfa1. Mechanical strain may act as a stimulator to induce differentiation of mesenchymal stem cells into osteoblasts, which is vital for bone formation in distraction osteogenesis.

Genetically engineered metal ion binding sites on the outside of a Channel's transmembrane beta-barrel.

We are exploring the ability of genetically engineered versions of the Staphylococcus aureus alpha-hemolysin (alphaHL) ion channel to serve as rationally designed sensor components for analytes including divalent cations. We show here that neither the hemolytic activity nor the single channel current of wild-type alphaHL was affected by [Zn(II)]

[Effects of Cistanche deserticola Y.C. Ma on serum creatine kinase and ultrastructures of skeletal muscles in mice].

The results showed that after mice were administrated with Cistanche deserticola, the duration of swimming was prolonged, and the increase of serum creative kinase was inhibited after exercises. Moreover, in the skeletal muscle ultrastructures it was found that after loaded swimming the glycogen became rich, and hyperplasia and hypertrophy of mitochondria occurred without any injury to myofibril.