4.4 kW Nd:YAG slab amplifier with a high repetition frequency and high beam quality.

Using the self-developed fused indium wetting technology and planar waveguide, the uniform heat dissipation of the slab crystal and uniform pumping of the pump light were achieved, respectively. Based on the master oscillator power amplification (MOPA) scheme, the power was then amplified when the seed light source passed through the Nd:YAG slab crystal three times. Additionally, the image transfer system that we added to the amplified optical path achieved high beam quality. Finally, we obtained a rectangular pulsed laser with an output average power of 4461 W, a repetition frequency of 20 kHz, a pulse width of 62 ns, an optical-to-optical conversion efficiency of 26.8%, and a beam quality of ß x=7.0 and ß y=7.7.

Dialogue between mitochondria and endoplasmic reticulum-potential therapeutic targets for age-related cardiovascular diseases.

Mitochondria-associated endoplasmic reticulum membranes (MAMs) act as physical membrane contact sites facilitating material exchange and signal transmission between mitochondria and endoplasmic reticulum (ER), thereby regulating processes such as Ca2+/lipid transport, mitochondrial dynamics, autophagy, ER stress, inflammation, and apoptosis, among other pathological mechanisms. Emerging evidence underscores the pivotal role of MAMs in cardiovascular diseases (CVDs), particularly in aging-related pathologies. Aging significantly influences the structure and function of the heart and the arterial system, possibly due to the accumulation of reactive oxygen species (ROS) resulting from reduced antioxidant capacity and the age-related decline in organelle function, including mitochondria. Therefore, this paper begins by describing the composition, structure, and function of MAMs, followed by an exploration of the degenerative changes in MAMs and the cardiovascular system during aging. Subsequently, it discusses the regulatory pathways and approaches targeting MAMs in aging-related CVDs, to provide novel treatment strategies for managing CVDs in aging populations.

UTILITY OF STREM-1 BIOMARKER AND HCP GENE FOR IDENTIFICATION OF ACINETOBACTER BAUMANNII COLONIZATION AND INFECTION IN LUNG.

ABSTRACT: Objective: Respiratory infections or colonization of Acinetobacter baumannii (Ab) are common in clinical practice but are treated differently. Early identification of Ab infection and colonization reduces the risk of antibiotic mismatch but objective laboratory indicators to distinguish between bacterial infections and colonization are lacking. To distinguish infection and colonization of Ab, we tested the role of two biomarkers, triggering receptor expressed on myeloid cells-1 (TREM-1) and hemolysin coregulated protein. Methods: A total of 96 inpatients with Ab were divided into infection and colonization groups. Blood samples were collected on days 1, 2, 3, 5, 8, and 10 and daily maximum body temperature was recorded. Polymerase Chain Reaction and Reverse Transcription Polymerase Chain Reaction were used to detect the presence and expression levels of the hcp gene in Ab clinical isolates. Results : sTREM-1 and procalcitonin (PCT) levels on days 1 to 10 and neutrophil classification (N%) on days 1 to 3 were different ( P < 0.05) in the infection group and colonization group. Receiver operating characteristic (ROC) curves showed significant differences in N% and sTREM-1 on days 2 and 3 ( P < 0.01). sTREM-1 had the highest AUC ROC on days 1, 2, and 3 of all the markers. On day 1, the ROC curve of "WBC&N%&PCT&sTREM-1" was statistically different from individual indices (white blood cell count, N%, and PCT; P < 0.05) and was equal to the ROC curve of sTREM-1 ( P > 0.05). Thirty five of 96 patients were classified as infection group and 61 as colonization group with hcp gene detection rates of 71.43% (25/35) and 31.15% (19/61), respectively. No differences in hcp gene presence and transcript levels were found between two groups ( P > 0.05). Conclusions: Dynamic monitoring of sTREM-1 and PCT is valuable in identifying Ab infection and colonization. sTREM-1 can be improved by combination with multiple biomarkers in the early stage for identification of infection and colonization. The hcp gene was more likely to be present in the infection cohort.

Genomic Determinants of Pathogenicity and Antimicrobial Resistance of Nosocomial Acinetobacter baumannii Clinical Isolates of Hospitalized Patients (2019-2021) from a Sentinel Hospital in Hangzhou, China.

Purpose: Acinetobacter baumannii (A. baumannii or AB) is one of the most opportunistic, nosocomial pathogens threatening public healthcare across countries. A. baumannii has become a primary growing concern due to its exceptional ability to acquire antimicrobial resistance (AMR) to multiple antimicrobial agents which is increasingly reported and more prevalent every year. Therefore, there is an urgent need to evaluate the AMR knowledge of A. baumannii for effective clinical treatment of nosocomial infections. This study aimed to investigate the clinical distribution AMR phenotypes and genotypes, and genomic characteristics of A. baumannii isolates recovered from hospitalized patients of different clinical departments of a sentinel hospital to improve clinical practices. Methods: A total of 123 clinical isolates were recovered from hospitalized patients of different clinical departments during 2019-2021 to analyze AMR patterns, and further subjected to whole-genome sequencing (WGS) investigations. Multi-locus sequence typing (MLST), as well as the presence of antimicrobial-resistant genes (ARGs), virulence factor genes (VFGs) and insertion sequences (ISs) were also investigated from WGS data. Results: The results highlighted that A. baumannii clinical isolates had shown a high AMR rate, particularly from the intensive care unit (ICU), towards routinely used antimicrobials, ie, ß-lactams and fluoroquinolones. ST2 was the most prevalent ST in the clinical isolates, it was strongly associated to the resistance of cephalosporins and carbapenems, with blaOXA-23 and blaOXA-66 being the most frequent determinants; moreover, high carrier rate of VFGs was also observed such as all strains containing the ompA, adeF, pgaC, lpsB, and bfmR genes. Conclusion: Acinetobacter baumannii clinical isolates are mostly ST2 with high rates of drug resistance and carrier of virulence factors. Therefore, it requires measurements to control its transmission and infection.

[Influence of the Ratio of Peripheral Hemoglobin-to-Red Cell Distribution width on the Prognosis of Patients with Diffuse Large B-cell Lymphoma].

OBJECTIVE: To investigate the influence of peripheral hemoglobin (Hb)-to-red cell distribution width (RDW) ratio (HRR) on the prognosis of patients with diffuse large B-cell lymphoma (DLBCL). METHODS: Data of 265 patients with diffuse large B-cell lymphoma (DLBCL) at the Affiliated Hospital of Xuzhou Medical University from January 2014 to December 2019 were retrospectively analyzed. 132 healthy people in the same period were used as normal control group. The best cut-off points of HRR was determined by receiver operating characteristics (ROC) curve; the chi-square test was used to analyze the correlation of clinical characteristics with HRR; the Kaplan-Meier method was used to compare the overall survival (OS) and progression-free survival (PFS) of HRR patients in different groups; the Cox proportional risk model was used for univariate and multivariate analysis. RESULTS: The best cut-off value of HRR was 0.936, which was divided into low HRR group and high HRR group. The low HRR group had a higher ECOG score, higher incidence of advanced Ann Arbor stage, higher NCCN-IPI score, and elevated LDH level. K-M survival analysis showed that OS (P<0.001) and PFS (P<0.001) in the low HRR group were significantly shorter than that in the higher HRR group. The multivariate analysis revealed that HRR was an independent predictor of OS（HR＝0.379，95％CI：0.237-0.605，P<0.001） and PFS （HR＝0.384，95％CI：0.241－0.614，P<0.001） in DLBCL patients. CONCLUSION: Low HRR（<0.936） in patients with DLBCL indicates a poor prognosis, which is an independent prognosis risk factor.

Understanding the influencing mechanism of users' participation in live streaming shopping: A socio-technical perspective.

Introduction: In live streaming shopping, exploring the influencing mechanism of consumers' participation is an important prerequi site for understanding consumer behavior in social commerce activities. The purpose of this study is to explore the relationship betw een technological and social factors (visibility, media richness, guidance shopping and real-time interactivity) in live streaming shop ping and consumers' purchase intention. The mediating roles of attraction and cognitive assimilation were also examined. Methods: This study collected 425 pieces of data through questionnaire survey. The structural equation model is established based on S-O-R frame. The hypothesis is tested by structural equation model. Results: Our study found that that real-time interactive and media richness positively affect attraction; visibility, guidance shopping and media richness positive affect cognitive assimilation; cognitive assimilation and attraction positive affect consumers' purchase intention; cognitive assimilation and attraction play a partial mediating role in the influence of technology and social factors on cons umers' purchase intention of live streaming shopping. Discussion: From the perspective of socio-technical, this study explores the influence mechanism of different influencing factors on consumers' purchase intention in live streaming shopping. This study expands the application of IT affordance theory in the context of live streaming shopping, and reveals the mediating role of attraction and cognitive assimilation between social, technological factors and consumers' purchase intention.

Treatment outcome of 301 aplastic anemia patients in China: a 10-year follow-up and real-world data from single institute experience.

OBJECTIVE: This study was carried out to explore clinical treatment and prognosis of patients with AA with different economic status. Methods: We retrospectively analyzed the clinical outcome of 301 patients with AA in our center from April 2008 to November 2017. RESULTS: Treatments included anti-thymocyte globulin (ATG) or anti-lymphocyte globulin (ALG) combined with cyclosporineA (CsA) (9%), allogeneic hematopoietic stem cell transplantation (allo-HSCT) (7%), CsA combined with androgen or CsA alone (hereinafter referred to as CsA group) (77%), no specific therapy (7%). The 5-year overall survival (OS) was higher in patients with non-severe AA (94.6%) compared with those with severe AA (SAA) (66.6%, P <.001), very severe AA (VSAA) (41.3%, P <.001). The 5-year OS was 76.5% in patients with SAA/VSAA treated with ATG/ALG combined with CsA, 75% in allo-HSCT group(P =.936), 63.6% in CsA group (P =.557), which was significantly higher than no specific therapy group (21.8%, P =.002). For those who responded to CsA , the duration of CsA (median follow-up time: 27 months, 1-101 months) was positively correlated with progression-free survival (r=0.603, P <.001). Multivariate analysis revealed that 36-65 years of age, SAA/VSAA, and no specific therapy were independent risk factors for inferior survival. CONCLUSION: The treatment of elderly patients with AA still faces challenges. CsA is benefit to the survival of SAA/VSAA patients. AA patients, who responded to initialy CsA treatment, may benefit from prolonged CsA treatment. In view of the side effects of CsA, the timing of withdrawal is worth further exploration.

Efficacy and safety of dapagliflozin in the treatment of chronic heart failure: A protocol for systematic review and meta-analysis.

BACKGROUND: As the last link in the chain of cardiovascular events, chronic heart failure (CHF) has high morbidity, high mortality, and poor prognosis. It is one of the main causes of death and disability worldwide. As a new drug for the treatment of chronic cardiovascular disease, dapagliflozin, the efficacy, and safety issues are still the focus of attention. Therefore, we conducted a meta-analysis to evaluate the efficacy and safety of dapagliflozin in the treatment of CHF. METHODS: According to the search strategy, regardless of publication date or language, randomized controlled trials (RCTs) of dapagliflozin for CHF will be retrieved from 8 databases. First of all, the literature was screened according to the eligibility criteria, and use the Cochrane Collaboration's tool to assess the quality of the included literature. Then, using Rev Man 5.3 and STATA 14.2 software for traditional meta-analysis. Finally, the evaluation of the quality of the evidence and the strength of the recommendations will adopt the Grading of Recommendations, Assessment, Development and Evaluation method. RESULTS: This study will evaluate the efficacy and safety of dapagliflozin for CHF, thereby providing more evidence support for clinical decision-making in CHF. CONCLUSION: Our research will provide more references for the clinical medication of patients with CHF. PROTOCOL REGISTRATION NUMBER: INPLASY202150046.

Study on the growth behavior and photoelectron spectroscopy of neodymium-doped silicon nanoclusters NdSin0/- (n = 8-20) with a double-hybrid density functional theory.

Structural evolution, magnetic moment, and thermochemical and spectral properties of NdSin0/- (n = 8-20) nanoclusters were studied. Optimized structures for NdSin demonstrated that the configuration with quintet ground state prefers Nd-substituted for a Si of the most stable Sin + 1 (n = 8-11) structure to Nd-linked configuration with Si9 tricapped trigonal prism subcluster (n = 12-19). Finally, the configuration prefers to Nd-encapsulated into Si cage framework (n = 20). For anion, the evolution at the quartet state prefers Nd-linked structure for n = 8-19 (excluded 9), and prefers Nd-encapsulated structure of n = 20. The spectral information including electron affinity, vertical detachment energy, and simulated photoelectron spectroscopy were also observed. The 4f electrons of Nd atom in NdSin with n = 8-10 hardly participate in bonding, but take part in remaining neutral clusters and all anionic NdSin- clusters. The calculations of average bond energy, HOMO-LUMO gap, and chemical bonding analyses reveal that NdSi20- possesses perfect thermodynamic and ideal chemical stability, making it as the most appropriate constitutional units for novel multi-functional semiconductors.

A drug-biomarker interaction model to predict the key targets of Scutellaria barbata D. Don in adverse-risk acute myeloid leukaemia.

A poor prognosis, relapse and resistance are burning issues during adverse-risk acute myeloid leukaemia (AML) treatment. As a natural medicine, Scutellaria barbata D. Don (SBD) has shown impressive antitumour activity in various cancers. Thus, SBD may become a potential drug in adverse-risk AML treatment. This study aimed to screen the key targets of SBD in adverse-risk AML using the drug-biomarker interaction model through bioinformatics and network pharmacology methods. First, the adverse-risk AML-related critical biomarkers and targets of SBD active ingredient were obtained from The Cancer Genome Atlas database and several pharmacophore matching databases. Next, the protein-protein interaction network was constructed, and topological analysis and pathway enrichment were used to screen key targets and main pathways of intervention of SBD in adverse-risk AML. Finally, molecular docking was implemented for key target verification. The results suggest that luteolin and quercetin are the main active components of SBD against adverse-risk AML, and affected drug resistance, apoptosis, immune regulation and angiogenesis through the core targets AKT1, MAPK1, IL6, EGFR, SRC, VEGFA and TP53. We hope the proposed drug-biomarker interaction model provides an effective strategy for the research and development of antitumour drugs.

Serum Small Proline-Rich Protein 2A (SPRR2A) Is a Noninvasive Biomarker in Gastric Cancer.

OBJECTIVE: Since early diagnosis is very important for treating gastric cancer (GC), we aimed to detect serum small proline-rich protein2A (SPRR2A) to verify its diagnostic value for GC patients. METHODS: Serum samples were collected from 200 patients with GC, 100 patients with gastritis, 40 patients with rectal cancer (RC), 50 patients with colon cancer (CC), and 100 healthy controls. An enzyme-linked immunosorbent assay (ELISA) detection kit was applied to measure serum SPRR2A concentration. The correlations between serum SPRR2A and carcinoembryonic antigen (CEA), clinical pathological parameters of GC, and receiver operating characteristic (ROC) curve were also analyzed. RESULTS: The median serum SPRR2A concentration in GC patients was significantly higher than those in healthy controls and gastritis or colorectal cancer patients (P < 0.001). Serum SPRR2A concentration at a cut-off value of 80.7 pg/ml yielded an AUC of 0.851, with 75.7% sensitivity and 74.5% specificity for discriminating GC patients from healthy people. The AUC for the serum SPRR2A concentration combined with the CEA concentration was 0.876, with 79.7% sensitivity and 78.7% specificity. Similarly, serum SPRR2A discriminated GC patients from gastritis patients with an AUC of 0.820, with 90.5% sensitivity and 61.7% specificity. The AUC for the serum SPRR2A concentration combined with the CEA concentration was 0.848, with 87.8% sensitivity and 68.1% specificity. The serum SPRR2A levels in GC patients were associated with lymph node metastasis and the tumor-node-metastasis (TNM) stage (P < 0.05). There was an obvious difference in serum SPRR2A expression between GC patients before and after surgery (P < 0.0001). CONCLUSION: These results suggest that serum SPRR2A can be used as an effective marker for GC.

A Novel Polymethyl Methacrylate Derivative Grafted with Cationic Iridium(III) Complex Units: Synthesis and Application in White Light-Emitting Diodes.

A novel orange-yellow-emitting polymethyl methacrylate derivative grafted with cationic iridium(III) complex units was synthesized and used as down-conversion luminescent materials in light-emitting diodes (LEDs). The polymer had a thermal decomposition temperature (Td) of 275 °C. With the temperature increasing from 20 to 100 °C, its photoluminescent intensity decreased to 76.8% with thermal quenching activation energy (Ea) of 0.2775 eV. A series of LEDs was fabricated by 460 nm blue GaN chips and the polymer blended in silicone at different concentrations. At 4.0 wt %, a cold white LED was obtained, the correlated color temperature (CCT) was 10,050 K, color rendering index (CRI) was 71.2, luminous efficiency (ηL) was 5.3 lm·w-1, and Commission Internationale de L'Eclairage (CIE) chromaticity coordinates were (0.30, 0.24). At 5.0 wt.%, the LED emitted neutral white light, its CCT was 4938 K, CRI was 75, ηL was 13.8 lm·w-1, and the CIE value was (0.34, 0.27). At 5.5 wt %, 6.0 wt %, 7.0 wt %, and 8.0 wt %, the LEDs all emitted warm white light; their CCTs were 3446, 3093, 2557, and 2337 K, respectively; their CRIs were 73.6, 71.8, 63.8, and 59.0, respectively; their ηL were 18.1, 16.3, 14.8, and 13.7 lm·w-1, respectively; and their CIE values were (0.36, 0.30), (0.40, 0.35), (0.45, 0.38), and (0.50, 0.42), respectively. At 9.0 wt %, the blue light of GaN chip was completely absorbed by the polymer and only the orange-yellow light of the polymer emitted. The results suggested the polymer was a promising orange-yellow-emitting phosphor candidate for white LEDs, especially for warm white LEDs.

Serum Exosomal Long Noncoding RNAs ENSG00000258332.1 and LINC00635 for the Diagnosis and Prognosis of Hepatocellular Carcinoma.

Background: Increasing studies suggest that long noncoding RNAs (lncRNAs) are involved in carcinogenesis of human cancers and might be used as diagnostic biomarkers for cancers.Methods: A total of 301 participants were recruited in the first part of the study, including a hepatocellular carcinoma (HCC) group (n = 60), liver cirrhosis (LC) group (n = 85), chronic hepatitis B (CHB) group (n = 96), and healthy subjects (n = 60). In the second part, we collected 55 HCC patients, 60 CHB patients, and 60 healthy subjects as an independent cohort to validate the ability of the experiential lncRNAs for identifying HCC from CHB. A commercial kit was used to isolate serum exosomes and total RNA. The relative levels of lnRNAs and GAPDH mRNA were measured with TaqMan PCR.Results: The results showed that the levels of ENSG00000258332.1 and LINC00635 in the HCC group were significantly higher than those in the other groups (all P < 0.05). A high ENSG00000258332.1 level in HCC was associated with portal vein tumor emboli, lymph node metastasis, TNM stage, and overall survival (OS; all P < 0.05), and a high LINC00635 level was related to lymph node metastasis, TNM stage, and OS (all P < 0.05). ENSG00000258332.1 discriminated HCC from CHB, gaining an area under the ROC curve (AUC) of 0.719 (cutoff value of 1.345); LINC00635 gained an AUC of 0.750 (cutoff value of 1.690). Furthermore, the AUC for the combination of the 2 lncRNAs and serum AFP (cutoff value of 20 µg/L) was 0.894. The abilities of the 2 lncRNAs for identifying HCC from CHB were validated by an independent cohort.Conclusions: The results suggested that the combination of serum exosomal ENSG00000258332.1, LINC00635, and AFP may be a valuable assay in diagnosis and prognosis of HCC.Impact: Our data will shed light on exosomal lncRNAs as biomarkers for HCC. Cancer Epidemiol Biomarkers Prev; 27(6); 710-6. ©2018 AACR.

Association between sn-2 fatty acid profiles of breast milk and development of the infant intestinal microbiome.

Increasing evidence shows that host diet and gut microbes are related. Previous studies have shown the effects of specific dietary fatty acids (FAs) on intestinal microbiota, but little is known about the effect of the stereospecifically numbered sn-2 position in triglycerides (TG) of human milk on the gut microbiome of infants. This study aimed at examining possible effects of sn-2 FAs of human milk on the gut microbial development of breastfeeding babies. Sn-2 FAs and intestinal microbiota were assessed by GC-MS and high-throughput 16S rRNA sequencing, respectively. The results showed that breast milk from mothers in China contained ten major sn-2 FAs dominated by palmitic acid (C16:0, 54.42%), oleic acid (C18:1 n-9, 14.95%), linoleic acid (LA, C18:2 n-6, 12.81%), myristic acid (C14:0, 4.50%) and C12:0 (3.17%). Total long chain unsaturated fatty acids (LCUFA) decreased from colostrum to mature milk, while total saturated fatty acids (SFA) showed no significant difference during lactation. A significant association between sn-2 FAs in milk and infant gut microbiota was found between decanoic acid (C10:0), myristic acid (C14:0), stearic acid (C18:0), C16:0, arachidonic acid (AA, C20:4 n-6), docosahexaenoic acid (DHA, C22:6 n-3) with Bacteroides, Enterobacteriaceae, Veillonella, Streptococcus, and Clostridium. These microbes were involved in short-chain fatty acid (SCFA) production and other functions, and significantly increased at 13-15 d after breastfeeding was initiated. C16:0 and DHA were relevant to most of the microbes. This study demonstrated the relatively steady profiles of sn-2 FAs in breast milk and gut microbiota of infants, together with their correlation during the breastfeeding period. The above results provided important information for designing the configuration of FAs in next-generation formulas for Chinese infants.

Serum exosomal hnRNPH1 mRNA as a novel marker for hepatocellular carcinoma.

BACKGROUND: Distinctive exosomal contents could be useful for cancer diagnosis and prognosis. However, little is known about whether serum exosomal heterogeneous nuclear ribonucleoprotein H1 (hnRNPH1) mRNA is a satisfactory biomarker for hepatocellular carcinoma (HCC). METHODS: Two hundred and ninety-one participants divided into four age- and gender-matched groups, including a HCC group (n=88), a liver cirrhosis (LC) group (n=67), a chronic hepatitis B (CHB) group (n=68) and a healthy control group (n=68), were enrolled. Serum exosomal hnRNPH1 mRNA and GAPDH mRNA were measured using TaqMan real-time PCR, and the relative expression levels were calculated. Receiver operating characteristic (ROC) curves were constructed to evaluate the effectiveness of hnRNPH1 mRNA alone and in combination with α-fetoprotein (AFP) in the diagnosis of HCC. The correlation between hnRNPH1 mRNA levels and clinicopathological characteristics and overall survival (OS) in HCC was determined. RESULTS: The serum exosomal hnRNPH1 mRNA levels in HCC patients were remarkably higher than in the other groups (p<0.05). The hnRNPH1 mRNA discriminated HCC from CHB with an area under the ROC curve (AUC) of 0.865, with sensitivity of 85.2% and specificity of 76.5% at cut-off value of 0.670. The AUC for hnRNPH1 mRNA in combination with AFP was further improved. The exosomal hnRNPH1 mRNA levels in HCC patients were associated with the Child-Pugh classification, portal vein tumor emboli, lymph node metastasis, TNM stage and OS (p<0.05). CONCLUSIONS: These findings suggested that serum exosomal hnRNPH1 mRNA could be an effective marker for HCC in high HBV prevalence areas.

Diagnostic Value of the Methylation of Multiple Gene Promoters in Serum in Hepatitis B Virus-Related Hepatocellular Carcinoma.

This study sought to evaluate the diagnostic value of the methylation of multiple gene promoters in serum in hepatitis B virus- (HBV-) related hepatocellular carcinoma (HCC). A total of 343 participants were enrolled, including 98 patients with HCC, 75 patients with liver cirrhosis (LC), 90 patients with chronic hepatitis B (CHB), and 80 healthy individuals. RASSF1A, APC, BVES, TIMP3, GSTP1, and HOXA9 were selected as the candidate genes. The MethyLight method was used to assay promoter methylation statuses. The diagnostic performances of markers were assessed by constructing receiver operating characteristic (ROC) curves. The prevalences of methylation for RASSF1A, APC, BVES, HOXA9, GSTP1, and TIMP3 were 52.04%, 36.73%, 29.59%, 20.41%, 17.35%, and 11.22%, respectively. APC methylation completely overlapped with RASSF1A methylation. The area under the curve (AUC) for RASSF1A methylation (0.718) was better than the corresponding AUC for AFP (0.609) in distinguishing HCC from CHB. When RASSF1A, BVES, HOXA9, and AFP were combined, the AUC was 0.852 (95% CI = 0.796-0.908, P = 0.028), and the sensitivity and specificity were 83.7% and 78.9%, respectively. In conclusion, an assay that combines methylation of the RASSF1A, BVES, and HOXA9 gene promoters in serum and AFP could significantly improve HBV-related HCC diagnoses.

Integrated DNA and RNA extraction using magnetic beads from viral pathogens causing acute respiratory infections.

Current extraction methods often extract DNA and RNA separately, and few methods are capable of co-extracting DNA and RNA from sputum. We established a nucleic acid co-extraction method from sputum based on magnetic beads and optimized the method by evaluating influencing factors, such as the guanidinium thiocyanate (GTC) and dithiothreitol (DTT) concentrations, magnetic bead amount, incubation temperature, lysis buffer pH and RNA carrier type. The feasibility of the simultaneous nucleic acid co-extraction method was evaluated by amplifying DNA and RNA viruses from a single clinical specimen with a multiplex RT-qPCR method. Both DNA and RNA were most efficiently extracted when the GTC and DTT concentrations were 2.0 M and 80 mM, respectively, 20 µl magnetic beads were added, the incubation temperature was 80 °C, the pH was 8 or 9, and RNA carrier A was used. Therefore, we established a simple method to extract nucleic acids from two important respiratory viruses compared with other commercial kits. This magnetic beads-based co-extraction method for sputum followed by a multiplex RT-qPCR can rapidly and precisely detect DNA and RNA viruses from a single clinical specimen and has many advantages, such as decreased time, low cost, and a lack of harmful chemicals.

Rapid and combined detection of Mycoplasma pneumoniae, Epstein-Barr virus and human cytomegalovirus using AllGlo quadruplex quantitative PCR.

Acute respiratory infections (ARIs) cause substantial morbidity and mortality worldwide. The causes of ARI are dynamic, and co-infections of Mycoplasma pneumoniae, Epstein-Barr virus and human cytomegalovirus are recently developed causes of ARI. Here, we established a quadruplex quantitative PCR (qPCR) method to rapidly identify and simultaneously detect a single infection or co-infection of these three pathogens and an internal control in a single tube using AllGlo probes. The analysis demonstrated a wide linear range of detection from 101 to 108 copies per test and a low coefficient of variation of less than 5 %. The amplification efficiencies were all close to 1, and the correlation coefficients (r2) were all greater than 0.99. We found no significant difference in a comparative reagent test (P >0.05). Moreover, the results of tests on clinical samples using AllGlo quadruplex qPCR and TaqMan uniplex qPCR were in near-perfect agreement (κ =0.97). Clinically, the availability of this method will enable better differential diagnosis, disease surveillance and controlled outcomes.

Combination of serum RASSF1A methylation and AFP is a promising non-invasive biomarker for HCC patient with chronic HBV infection.

BACKGROUND: Hypermethylation of the promoter region of the RAS association domain family 1A gene (RASSF1A) occurs widely in hepatocellular carcinoma (HCC) tissues. While the diagnostic performance of the use of RASSF1A methylation as a serum or plasma marker in patients with HCC has varied largely in the literature,we confirmed the clinical application value of serum RASSF1A methylation for HBV related HCC in this study. METHODS: A total of 584 participants were recruited into this study, including 190 patients with HCC, 114 patients with liver cirrhosis (LC), 120 patients with chronic hepatitis B (CHB) and 160 healthy individuals. Serum RASSF1A methylation was determined by the MethyLight method. In addition, we followed up 43 HCC patients who were unable to undergo surgery for 24 months. RESULTS: Serum RASSF1A methylation occurred significantly more frequently in patients with HCC (122/190, 64.2%) than in patients with LC (20/114, 17.5%), patients with CHB (6/120, 5.0%) and in healthy individuals (0/160, 0) (P < 0.001); moreover, it allowed for the discrimination of patients with HCC from those with CHB with an areas under the ROC curves (AUC) of 0.796 (64.2% sensitivity and 89.8% specificity). Furthermore, the AUC for the combination of serum RASSF1A methylation and AFP level (≥20 ng/L) was 0.876 (80.9% sensitivity and 93.4% specificity). Serum RASSF1A methylation positive in patients with HCC was associated with more malignant clinical characteristics and a worse overall survival (OS) (P < 0.05). CONCLUSION: Serum RASSF1A methylation demonstrated a satisfactory value for in the diagnosis of HBV related HCC, and could predict clinical progression and prognosis. In addition, our findings suggested that the combination of serum RASSF1A methylation and AFP level may be a promising non-invasive biomarker for the discrimination of patients with HCC from those with CHB. VIRTUAL SLIDES: The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_DPAT-D-15-00090.1.

Serum miR-96 is a promising biomarker for hepatocellular carcinoma in patients with chronic hepatitis B virus infection.

MicroRNAs (miRNAs) are involved in cancer biology, and some distinctive serum miRNAs could be useful for cancer diagnosis and prognosis. However, little is known about whether serum miR-96 is a satisfactory biomarker for hepatocellular carcinoma (HCC). Four hundreds and fourteen participants were enrolled in this study, and they were divided into four age- and gender-matched groups, including the HCC group (n = 104), liver cirrhosis (LC) group (n = 90), chronic hepatitis B (CHB) group (n = 100) and healthy control group (n = 120). Serum miR-96 was measured by real-time PCR, the levels of which were calculated by the 2(-ΔCt) method. Serum miR-96 levels in the HCC patients were remarkably higher than in the other groups (P < 0.01), and the serum miR-96 levels discriminated HCC patients from CHB patients with an area under the ROC curve (AUC) of 0.803 (77.9% sensitivity and 75.3% specificity). Furthermore, the AUC for combined miR-96 and α-fetoprotein (AFP) was 0.889 (83.6% sensitivity and 82.4% specificity). High serum miR-96 levels in HCC patients were associated with larger tumor size, higher prevalence of lymph node metastasis, higher TNM stage and worse overall survival (OS) (P < 0.05). Our findings suggest that serum miR-96 is a promising biomarker for HCC patients with chronic HBV infection.