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A unique and versatile ratiometric electrochemical DNA biosensor was designed for the first time by integration of homogeneous exonuclease III-assisted target recycling amplification and one-step triggered dual-signal output. The presence of the target DNA would hybridize with the specifically designed ferrocene-labeled hairpin probe (Fc-P1) and initiate homogeneous exonuclease III-assisted target recycling amplification, which led to the decrease of the local concentration of Fc-P1. The target DNA dependent amount of remaining Fc-P1 could be conveniently monitored by hybridize the remaining Fc-P1 with the methylene blue-labeled hairpin probe ((MB-PP1) on the sensor electrode to trigger the MB tags away from while the Fc tags close to the sensing interface. Therefore, such recognition event can be translated into the homogeneous amplification coupled with one-step triggered dual-signal ratiometric electrochemical readout. The proposed biosensor achieves a detection limit of 12.8 fM within a linear range of 0.02 pM to 2 nM for target DNA species related to oral cancer over expressed 1, along with a favorable specificity. More importantly, the biosensor had been applied in detection of target DNA in artificial saliva sample and provided significant potential application in clinical analysis. This strategy possessed good capacity to integrate the nuclease amplification techniques with ratiometric output modes, and would open new opportunities for sensitive detection of other biorecognition events. [ABSTRACT FROM AUTHOR]

A simple and novel one-step triggered ratiometric electrochemical biosensor was designed for lead (Pb 2+ ) based on the highly specific lead-dependent DNAzyme and dual-signal output mode. The biosensor consists of a thiolated methylene blue-labeled DNA (MB-P1) as catalytic probe and “signal-on” readout, and a complementary strand modified with ferrocene (Fc-P2) as substrate probe and “signal-off” output. The presence of Pb 2+ could activate the DNAzyme and cleave the sessile phosphodiester of the Fc-P2 into two fragments, which lead to the departure of Fc from the sensing interface along with the single stranded MB-P1 near the electrode surface. Therefore, the Pb 2+ recognition event resulted in both the “signal-on” of MB and the “signal-off” of Fc for dual-signal ratiometric electrochemical readout. Combined with the efficient recognition capacity of the designed DNAzyme and the dual-signal amplification strategy, the proposed biosensor showed a wide detection range from 0.1 nM to 5 μM with a detection limit of 45.8 pM (S/N = 3). Meanwhile, this DNAzyme-based Pb 2+ biosensor exhibits reasonable selectivity, fast analytical speed, acceptable fabrication reproducibility, and operational convenience. More importantly, the system is capable of detecting Pb 2+ in biological fluid such as serum, suggesting promising applications of this biosensor in on-site and real-time clinical Pb 2+ detection. [ABSTRACT FROM AUTHOR]

Background Early diagnosis of hepatitis C virus (HCV) infection is essential to allow appropriate treatment and prevent transmission. Objectives To evaluate the Elecsys ® Anti-HCV II assay as a routine screening assay in Asia using a large number of samples from different Asian Pacific populations and compare its performance with other HCV assays routinely used in the region. Study design The sensitivity and specificity of the Elecsys ® Anti-HCV II assay were determined using routine hospital samples and compared with at least one of the following comparator assays at nine independent centers: ARCHITECT™ Anti-HCV; Serodia ® -HCV Particle Agglutination; Vitros ® ECi Anti-HCV; Elecsys ® Anti-HCV; ADVIA Centaur ® HCV; InTec ® HCV EIA; or Livzon ® Anti-HCV. Commercially available seroconversion panels were used to assess sensitivity for early detection of infection. Results The Elecsys ® Anti-HCV II assay was more sensitive in recognizing early infection and detected acute HCV infection earlier on average than the comparator assays for all six panels tested. 7,726 routine samples were tested and 322 identified as HCV positive. Elecsys ® Anti-HCV II had a sensitivity of 100% and a specificity of 99.66%, both of which were comparable or superior to the results obtained for competitor assays, which ranged from 87.5–100% and 98.98–100%, respectively. Conclusions The Elecsys ® Anti-HCV II assay has the sensitivity and specificity to support its use as a routine screening method in the Asia Pacific region. Furthermore, this assay shortens the diagnostic window between infection and the detection of antibodies compared with established methods. [ABSTRACT FROM AUTHOR]

Background: Long non-coding RNAs (LncRNAs) have emerged as important regulators in many physiological processes in plant. By high-throughput RNA-sequencing, many pathogen-associated LncRNAs were mapped in various plants, and some of them were proved to be involved in plant defense responses. The rice blast disease caused by Magnaporthe oryzae (M. oryzae) is one of the most destructive diseases in rice. However, M. oryzae-induced LncRNAs in rice is yet to be studied. Findings: We investigated rice LncRNAs that were associated with the rice blast fungus. Totally 83 LncRNAs were up-regulated after blast fungus infection and 78 were down-regulated. Of them, the natural antisense transcripts (NATs) were the most abundant. The expression of some LncRNAs has similar pattern with their host genes or neighboring genes, suggesting a cis function of them in regulating gene transcription level. The deferentially expressed (DE) LncRNAs and genes co-expression analysis revealed some LncRNAs were associated with genes known to be involved in pathogen resistance, and these genes were enriched in terpenoid biosynthesis and defense response by Gene Ontology (GO) enrichment analysis. Interestingly, one of up-regulated DE-intronic RNA was derived from a jasmonate (JA) biosynthetic gene, lipoxygenase RLL (LOX-RLL). Levels of JAs were significantly increased after blast fungus infection. Given that JA is known to regulate blast resistance in rice, we suggested that LncRNA may be involved in JA-mediated rice resistance to blast fungus. Conclusions: This study identified blast fungus-responsive LncRNAs in rice, which provides another layer of candidates that regulate rice and blast fungus interactions. [ABSTRACT FROM AUTHOR]

A dual‐polarized wide‐beam (DPWB) antenna is optimized by a hybrid topology optimization (HTO) method, which combines the scalar isotropic material with penalization and the level set method. The HTO method has high optimization freedom degree, strong topology‐searching ability and fast convergence speed. The initial model of the antenna is designed as a dual‐feeding square microstrip patch antenna with short‐circuit metal walls around the patch. By optimizing the metal patch on the upper side of the model, the DPWB antenna is obtained. The DPWB antenna is symmetric with respect to both ports. When fed to either port, the half‐power beamwidth of the DPWB antenna can reach 166° (−82° ~ 84°) in the E‐plane and 124° (−62° ~ 62°) in the H‐plane. The performance is verified by both simulation and measurement. The DPWB antenna is further applied to a linear phased array, reaching an E‐plane scanning angle range of 160° (−80° ~ 80°) and an H‐plane scanning angle range of 122° (−61° ~ 61°), with the gain fluctuation less than 3 dB and the sidelobe level less than −10.1 dB. [ABSTRACT FROM AUTHOR]

Background: Preeclampsia (PE) is relatively common and is unpredictable in its onset and progression. Aims: We investigated the clinical value of using the multiple of the median (MoM) of free β-human chorionic gonadotropin (β-hCG) concentrations in women with normal pregnancy and PE. Methods: This study was based on a dataset available from published studies, and the relevant studies were retrieved from multiple electronic databases. Data were extracted from case-control studies; a random-effects model was employed, and standardized mean difference and 95% confidence intervals were calculated. Twelve case-control studies (eleven English-based articles and one Chinese-based article) were analyzed in the current meta-analysis and included 702 patients with PE and 8,233 women with normal pregnancies. Results: Statistical analysis revealed a higher MoM of β-hCG serum levels in patients with PE. Ethnicity subgroup analysis showed that the MoM of serum β-hCG levels was significantly higher in women with PE in both Asian and Caucasian populations. Conclusion: The MoM of β-hCG serum levels was significantly increased in women with PE compared to women with normal pregnancies. Screening for serum β-hCG MoM levels will be helpful in the early identification of pregnancies at risk of developing PE. [ABSTRACT FROM AUTHOR]

Objective To explore the the correlations of p16INK4A (p16) and survivin expressions with cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma. Methods The p16 and survivin expressions were detected in 50 cervical squamous cell carcinoma tissues, 150 various grades of CIN tissues and 30 normal cervical tissues using immunohistochemistry. All data were analyzed applying SPSS 17.0 software. Results The p16 and survivin expressions showed the presence of statistical significance in cervical cancer, CINI, CINII, CINIII and normal cervical tissues ( P < 0.05), and the comparison also revealed statistical significance among groups (all P < 0.05); the p16 and survivin expressions were positively correlated with the grade of cervical diseases (both P < 0.05). Moreover, p16 protein was associated with CIN grade and lymph node metastases in cervical cancer (all P < 0.05); survivin protein was also related with clinical stages, CIN grade and lymph node metastases (all P < 0.05); the p16 and survivin expressions were positively correlated with cervical cancer (r = 0.854, P < 0.001), and associated with poor prognosis of cervical cancer. Conclusion Briefly, p16 and survivin expression may be correlated with the clinico-pathological and prognosis of cervical cancer. [ABSTRACT FROM AUTHOR]

Kidney transplantation is the ideal treatment for end-stage renal disease (ESRD). Chronic antibody-mediated rejection (CAMR) is the main cause of graft failure. Tfh and B cells are key immune cells that play important roles in CAMR. In this study, the populations of different Tfh cell phenotypes and B cell subsets in CAMR were investigated in a total of 36 patients. Based on Banff-2019, 15 patients were diagnosed with CAMR (CAMR group), 11 recipients were diagnosed with recurrent or de novo IgA nephropathy (IgAN group), and 10 patients displayed stable renal function (stable group). The Tfh and B cell subsets were analyzed by flow cytometry. The percentage and absolute number of PD-1+ICOS+Tfh cells were significantly higher in CAMR (p < 0.05), as was the ratio of CD226+Tfh cells to TIGIT+Tfh cells (p < 0.05). Compared with stable recipients, CAMR patients had lower naïve B cells and higher unswitched memory B cells, which were also significantly related to renal function (p < 0.05). Using the logistic regression model, we concluded that the estimated glomerular filtration rate (eGFR), absolute number of PD-1+ICOS+Tfh cells, and ratio of CD226+Tfh cells to TIGIT+Tfh cells were independent risk factors for CAMR. The combination of eGFR, PD-1+ICOS+Tfh cells, and the ratio of CD226+Tfh cells to TIGIT+Tfh cells showed better diagnostic efficacy for CAMR than each single parameter. The collective findings show that monitoring different Tfh phenotypes and B cell subsets is beneficial to kidney transplant recipients and implicate the combination of eGFR, number of PD-1+ICOS+Tfh cells, and ratio of CD226+Tfh cells to TIGIT+Tfh cells as a biomarker for diagnosing CAMR. The findings may also inform new strategies to identify and treat CAMR. [ABSTRACT FROM AUTHOR]

Summary: Background: Current guidelines recommend echinocandins as first‐line therapy for candidemia. However, several non‐Candida yeast are non‐susceptible to echinocandins (echinocandin non‐susceptible yeast, ENSY), including Cryptococcus, Geotrichum, Malassezia, Pseudozyma, Rhodotorula, Saprochaete, Sporobolomyces and Trichosporon. In laboratories that are not equipped with rapid diagnostic tools, it often takes several days to identify yeast, and this may lead to inappropriate presumptive use of echinocandins in patients with ENSY fungemia. The aim of this study was to determine the distribution of ENSY species during a 1‐year, laboratory surveillance programme in Asia. Methods: Non‐duplicate yeast isolated from blood or bone marrow cultures at 25 hospitals in China, Hong Kong, India, Singapore, Taiwan and Thailand were analysed. Isolates were considered to be duplicative if they were obtained within 7 days from the same patient. Results: Of 2155 yeast isolates evaluated, 175 (8.1%) were non‐Candida yeast. The majority of non‐Candida yeast were ENSY (146/175, 83.4%). These included Cryptococcus (109 isolates), Trichosporon (23), Rhodotorula (10) and Malassezia (4). The proportion of ENSY isolates (146/2155, 6.7%) differed between tropical (India, Thailand and Singapore; 51/593, 8.6%) and non‐tropical countries/regions (China, Hong Kong and Taiwan; 95/1562, 6.1%, P = 0.038). ENSY was common in outpatient clinics (25.0%) and emergency departments (17.8%) but rare in intensive care units (4.7%) and in haematology‐oncology units (2.9%). Cryptococcus accounted for the majority of the non‐Candida species in emergency departments (21/24, 87.5%) and outpatient clinics (4/5, 80.0%). Conclusions: Isolation of non‐Candida yeast from blood cultures was not rare, and the frequency varied among medical units and countries. [ABSTRACT FROM AUTHOR]

Background T follicular helper cells (Tfh) are recently revealed to be vital in antibody-mediated rejection (AMR) in kidney transplant recipients (KTRs). However, the impact of immunosuppressive drugs on Tfh cells is not fully understood. The purpose of this study was to investigate the variation of Tfh cells phenotypically and functionally in KTRs treated with different immunosuppression regimens. Methods We recruited 26 KTRs treated with tacrolimus (TAC) -based regimen, 13 with sirolimus (SRL) -based regimen and 10 healthy controls (HC) in this study. The percentage and absolute number of circulating Tfh cells and the co-expression of Tfh related molecules including inducible costimulatory molecule (ICOS), programmed cell death protein 1 (PD-1), interleukin-21 (IL-21) and signal transducer and activator of transcription 3 (STAT3) were analyzed by flow cytometry, while serum IL-6 was detected by electrochemiluminescence immunoassay. Results The percentage and absolute number of Tfh cells and the co-expression of PD-1, STAT3 in Tfh cells were significantly higher in TAC group than that in SRL group. While no difference was found in regard to IL-21 and ICOS co-expressed with Tfh cells among three groups. Multiple linear regression analysis results showed that pre-transplant PRA level was the significant confounder affecting the absolute numbers of Tfh and CD4 + CXCR5 + PD-1 + T cells. In addition, correlation analysis showed that CD4 + CXCR5 + STAT3 + T cells were positively correlated to Tfh cells. Conclusions Our study indicates that sirolimus can suppress the quantity of Tfh cells more significantly than tacrolimus. The higher level of circulating Tfh cells in tacrolimus group might be related to STAT3 signaling. [ABSTRACT FROM AUTHOR]

Background T cell immunoglobulin mucin-3 (Tim-3) has been reported to participate in the regulation of immune response and the induction of allograft tolerance. However, the association between Tim-3 and renal allograft dysfunction is unclear. We studied the expression of cellular and soluble Tim-3 (sTim-3), soluble galectin-9 (sGal-9) and carcinoembryonic antigen-related cell adhesion molecule-1 (sCEACAM-1) in kidney transplantation recipients (KTRs) to explore their roles in allograft dysfunction. Methods 96 KTRs (53 with stable graft and 43 with graft dysfunction) and 30 healthy controls (HC) were enrolled. Among the KTRs, 55 used Tacrolimus (TAC) and 41 used Sirolimus (SRL). In the dysfunction group, 29 recipients have undergone graft biopsy and 14 were classified as biopsy-proven rejection (BPR). Cellular Tim-3 was determined by flow cytometry. sTim-3 was determined by ELISA. sGal-9 and sCEACAM-1 were determined by Bio-Plex® suspension array system. Results KTRs with renal dysfunction showed significantly higher levels of sTim-3 and sGal-9 but similar levels of cellular Tim-3 and sCEACAM-1 compared with stable recipients. Correlation analysis revealed that estimated glomerular filtration rate (eGFR) was negatively associated with sTim-3 and sGal-9. Both BPR and non-BPR groups showed comparable levels of Tim-3, Gal-9 and CEACAM-1. Moreover, SRL group showed significantly higher levels of sCEACAM-1 than TAC and HC groups. Conclusions sTim-3 and sGal-9 were promising biomarkers for allograft dysfunction, but unable to differentiate allograft rejection from other causes of renal dysfunction in KTRs. Moreover, long-term administration of sirolimus would up-regulate sCEACAM-1 level, while exert similar regulatory effects on Tim-3 and Gal-9 compared to tacrolimus. [ABSTRACT FROM AUTHOR]

Aims Elderly transplant recipients have a lower incidence of acute rejection, and a higher risk to die from infectious complications. A potential cause may be differences in the pharmacokinetics (PK) or pharmacodynamics (PD) of the immunosuppressive drugs they are taking. This study was designed to comprehensively evaluate the influence of age on the PK and PD of mycophenolic acid (MPA). Methods In this study the PK and PD of MPA was studied in 26 elderly and 54 younger renal transplant recipients treated with mycophenolate mofetil and tacrolimus. Patients were sampled repetitively, both before and during the first 6 months after kidney transplantation. Age-related variability in MPA PK, baseline IMPDH activity, as well as MPA-induced IMPDH inhibition were studied. Results The IMPDH activity pre-transplantation did not differ between elderly and younger patients. Neither IMPDH activity pre-transplantation nor maximum IMPDH inhibition was significantly correlated with the patients' age. The area under the MPA plasma concentration-time curve (AUC0-12h) and the area under the effect (IMPDH activity)-time curve (AEC0-12h) from 0 to 12 h were also not significantly different between the two groups. We found no significant differences in EC50 and Emax between elderly and younger patients. Conclusions Age did not significantly affect the PK or PD of MPA. It is unlikely that the lower incidence of acute rejection in elderly patients, or the higher risk to die from a severe infection in elderly patients is due to different handling of MPA in the elderly. [ABSTRACT FROM AUTHOR]

Objective To establish the CTRP4 transgenic mouse model and investigate the function of the novel adipocytokine CTRP4. Methods CTRP4 overexpressing vector in pCAGGS was firstly constructed and then microinjected into zygote to establish the founder transgenic mice. Fl heterozygotes were generated by founder mice mating with wildtype mice, and the CTKP4 transgenic homozygotes were generated by F 1 littermates. The genotype was confirmed by PCR and test cross method. The expression level of CTKP4 in transgenic mice was delected by western blot. Result The human CTRP4 transgenic homozygote mice line was established, and the expression level of CTRP4 was confirmed raletively high in detected tissues including heart, liver, brain and kidney. Conclusion The human CTRP4 transgenic mice was successfully established. [ABSTRACT FROM AUTHOR]

Background: Acute kidney injury (AKI) has been proposed as a leading cause of mortality for acute pancreatitis (AP) patients admitted to the intensive care unit (ICU). This study investigated the predictive value of procalcitonin (PCT) for AKI development and relevant prognosis in patients with AP, and compared PCT’s predictive power with that of other inflammation-related variables. Methods: Between January 2011 and March 2013, we enrolled 305 cases with acute pancreatitis admitted to ICU. Serum levels of PCT, serum amyloid A (SAA), interleukin-6 (IL-6), and C reactive protein (CRP) were determined on admission. Serum PCT was tested in patients who developed AKI on the day of AKI occurrence and on either day 28 after occurrence (for survivors) or on the day of death (for those who died within 28 days). Results: Serum PCT levels were 100-fold higher in the AKI group than in the non-AKI group on the day of ICU admission (p<0.05). The area under the receiver-operating characteristic (ROC) curve of PCT for predicting AKI was 0.986, which was superior to SAA, CRP, and IL-6 (p<0.05). ROC analysis revealed all variables tested had lower predictive performance for AKI prognosis. The average serum PCT level on day 28 (2.67 (0.89, 7.99) ng/ml) was significantly (p<0.0001) lower than on the day of AKI occurrence (43.71 (19.24,65.69) ng/ml) in survivors, but the serum PCT level on death (63.73 (34.22,94.30) ng/ml) was higher than on the day of AKI occurrence (37.55 (18.70,74.12) ng/ml) in non-survivors, although there was no significant difference between the two days in the latter group (p = 0.1365). Conclusion: Serum PCT is superior to CRP, IL-6, and SAA for predicting the development of AKI in patients with AP, and also can be used for dynamic evaluation of AKI prognosis. [ABSTRACT FROM AUTHOR]

Background: The Elecsys® HIV combi PT assay was developed to allow earlier detection of HIV infection with increased sensitivity and specificity. Objectives: To validate the assay for screening and reliable early detection of HIV-1 infection in Asia. Study design: Samples tested reflected those routinely screened in Asia and comprised: HIV-1 antigen lysate (25 samples) and antibody (20 samples) dilutions; seven HIV-1 seroconversion panels (46 samples); 39 patient samples from early infection; 183 known-positive sera; HIV-1 p24 antigen sensitivity panel (seven samples); >500 routine clinical samples per center. The Elecsys® HIV combi PT assay was compared with fourth- (ADVIA Centaur® HIV combo, ARCHITECT® HIV combo, Elecsys® HIV combi) and third-generation (VIRONOSTIKA® HIV Uni-Form II Plus O, Zhuhai Livzon Anti-HIV EIA, Serodia® Particle Agglutination) assays commonly used in the region. Results: Overall, the Elecsys® HIV combi PT showed superior or similar sensitivity to the comparators for detecting all subtypes. The assay correctly identified all positive samples, including those taken soon after infection, and detected seroconversion at a similar or shorter time interval than the comparators. The analytical sensitivity of Elecsys® HIV combi PT for HIV-1 p24 antigen was 0.90IU/mL, which was lower than reported previously. The assay showed good specificity (99.86%) that was superior or equivalent to the other fourth-generation assays tested. Conclusions: These robust data demonstrate the good subtype inclusivity of the Elecsys® HIV combi PT assay and its suitability for screening and reliable early detection of HIV infection in Asia. [ABSTRACT FROM AUTHOR]

Proinflammatory Th17 cells and CD4CD25 regulatory T (Treg) cells are two newly identified T lymphocyte subsets, which have opposite effects on autoimmunity and inflammation. To assess the Th17/Treg pattern and cytokine microenvironment in peripheral blood of patients with RA, we included 66 RA patients and 20 healthy volunteers. Of all these subjects, peripheral Th17 and Treg frequencies were analyzed by flow cytometry (FCM) and the plasma levels of interleukin (IL)-17, 23, 6, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1 were detected by ELISA. The results demonstrated that RA patients revealed an obvious increase in peripheral Th17 frequencies and levels of Th17-related cytokines (IL-17, IL-23, IL-6, TNF-α) while a significant decrease in Treg frequencies and Treg-related cytokine (TGF-β1) levels when compared with healthy people. Our study indicated that development of RA is associated with peripheral Th17/Treg imbalance and characterized by a proinflammatory cytokine microenvironment, which supports continuing generation of Th17 cells. [ABSTRACT FROM AUTHOR]

Objective: To compare the perioperative systemic immunity of laparoscopy-assisted and open radical gastrectomy for gastric cancer. Methods: Patients with gastric adenocarcinoma proven by endoscopy and biopsy were eligible, while patients with preoperative staging of T4, N2-3, or M1 were excluded. Eligible patients willing to undertake laparoscopic surgery in the consecutive cohort were assigned to the laparoscopy-assisted gastrectomy (LAG) group, while concurrent patients were assigned to the conventional open gastrectomy (OG) group. All operations were performed with the intention of radical resection. Various immunological parameters were tested in peripheral venous blood collected at preoperative 1st day and postoperative 2nd day (POD2) and 7th day (POD7). SPSS 13.0 software was used for statistical analysis. Results: Thirty patients were included, 15 each in the LAG and OG groups. The general characteristics and short-term outcomes (harvested lymph nodes number, hospital stay, complications, and mortality rate) of the two groups were comparable, but the operation time was significantly longer in LAG (P = 0.001). Moreover, intergroup comparisons indicated no significant differences between the groups in levels of neutrophils, T-lymphocytes, natural killer cells, IgG, IgM, IgA, C3, C4, interleukin-6, or interleukin-10 at any time point (P > 0.05). However, there was a gradual decrease in natural killer cell count in the LAG group up to POD7 (P = 0.008). Conclusion: The changes in systemic immunity markers were comparable between laparoscopy-assisted and open gastrectomy for gastric cancer. However, there was a trend of suppression of natural killer cells in the laparoscopy-assisted gastrectomy group. [ABSTRACT FROM AUTHOR]

Screening for hepatitis B surface antigen (HBsAg) demands highly sensitive and specific immunoassays with the ability to detect clinically relevant surface gene mutants—the presence of which is an increasing concern and can compromise test results. The objective of the study was to compare the clinical and technical performance of the fully automated Elecsys HBsAg II assay with other widely used HBsAg immunoassays in China. This was a multicentre study in which eight Chinese laboratories compared the Elecsys HBsAg II assay with the Architect HBsAg assay, AxSYM HBsAg assay, or generic microtitre plate (MTP) enzyme-linked immunosorbent assays (manufactured in China) against preselected samples, including recombinant surface gene mutants, and routine clinical samples. Elecsys HBsAg II was the most sensitive assay for detecting positive results in seroconversion samples: up to 14, 11, and 22 days earlier than the Architect, AxSYM, and MTP HBsAg assays, respectively. It also detected all 211 preselected HBsAg-positive specimens and all 13 recombinant HBsAg mutants; the AxSYM and MTP HBsAg assays failed to detect 3 and 10 mutant samples, respectively. Sensitivity was 100% for Elecsys HBsAg II with routine samples, compared with 99.1, 98.9, and 95.2% for the Architect, AxSYM, and MTP HBsAg assays, respectively. In conclusion, it was demonstrated that the Elecsys HBsAg II assay is suitable for routine HBsAg screening in China. [ABSTRACT FROM AUTHOR]

Here we show that fusion of two complementarity-determining regions (CDRs), VHCDR1 and VLCDR3, through a cognate framework region (VHFR2) yields mimetics that retain the antigen recognition of their parent molecules, but have a superior capacity to penetrate tumors. The antigen-recognition abilities of these ∼3 kDa mimetics surpass those of comparable fragments lacking the framework region. In vivo activities of the mimetics suggests that the structural orientation of their CDRs approximates the conformation of the CDRs in the complex of the parent antibody with antigen. We linked the antibody mimetics to the bacterial toxin colicin Ia to create fusion proteins called “pheromonicins,” which enable targeted inhibition of tumor growth. In mice bearing human malignant tumors, pheromonicins directed against tumor-specific surface markers show greater capacity to target and penetrate tumors than their parent antibodies. Rational recombination of selected VH/VL binding sites and their framework regions might provide useful targeting moieties for cytotoxic cancer therapies. [ABSTRACT FROM AUTHOR]

• A sensitive UPLC–MS/MS method is developed for azathioprine metabolites in blood. • The method displayed satisfactory validation parameters. • Using a simple and fast intracellular concentration normalization algorithm. • First reporting the concentrations of azathioprine metabolites in AIH patients. Individualized therapy involves genetic test of drug metabolism, which provides information about the initial dose and therapeutic drug monitoring for adjusting the subsequent dose. Consequently, toxic side effects are expected to be minimized and therapeutic effects to be maximized. In this study, an ultra-performance liquid chromatography tandem mass spectrometry method that was specific, accurate and sensitive was developed to simultaneously determine azathioprine two metabolites, 6-thioguanine nucleotides (6-TGN) and 6-methyl-mercaptopurine riboside (6-MMPr) in the whole blood lysate. We precipitated the sample by trifluoroacetic acid under the protection of dithiothreitol, with 6-MMPr and 6-TGN being hydrolyzed to produce 6-methymercaptopurine and 6-thioguanine. In the chromatographic separation, Waters ACQUITY BEH C18 (2.1 × 100 mm, 1.7 μm) chromatographic column was applied and gradient elution was conducted with 0.02 mol/L ammonium acetate buffer (which contains 0.3% formic acid) and acetonitrile at a flow rate of 0.4 ml/min. Tandem mass spectrometry in multiple reaction monitoring mode was applied for detection via electrospray ionization source in positive ionization mode. The analyzing process lasted for no more than 2 min. The calibration curve for each metabolite fitted a least squares model (weighed 1/X) from 1.25 to 5000 ng/ml (r2 > 0.99). The ion pairs were detected as 6-MMP m / z 167.07 → 152.15, 6-TG m / z 168.06 → 134.13, and internal standard m / z 171.07 → 137.14. Under the guidance of FDA guidelines for bioanalytical method validation, we carried out validation and obtained satisfactory results. The method was successfully utilized for monitoring the concentrations of each metabolite from 65 affected patients who had received azathioprine maintenance therapy and achieved optimal results. [ABSTRACT FROM AUTHOR]

Infection remains a major cause of morbidity and mortality after kidney transplantation (KT). Reliable biomarkers to predict post-transplant infection are lacking. We investigated the predictive performance of pre- and post-transplant levels of T-cell immunoglobulin and mucin domain-3 (Tim-3) and Galectin-9 (Gal-9), two pleiotropic immunomodulatory molecules, in early identification of infection. Serum Tim-3 and Gal-9 were paired measured before and 30 days after transplantation (PTD 30) in 95 KT recipients (KTRs). The decline rates of Tim-3 and Gal-9 were calculated relative to pre-transplant levels. KTRs with infection history had significantly higher levels of PTD 30 Tim-3 and Gal-9, and slower decrease rates of Gal-9 compared to non-infected recipients, while no difference was observed between two groups regarding pre-transplant levels. The AUCs for predicting 1-year post-transplant infection were 0.653 and 0.711 for post-transplant Tim-3 and Gal-9, 0.664 and 0.670 for relative Tim-3 and Gal-9, respectively. After adjusting for potential confounders, PTD 30 Tim-3, Gal-9 and relative Gal-9 remained as independent risk factors for post-transplant infection. Our results suggested that PTD 30 Tim-3 and Gal-9 and relative decrease of Gal-9 were promising predictors for identifying KTRs with high risk of infection, while pre-transplant Tim-3 and Gal-9 showed no predictive power to infection. • Higher serum levels of PTD 30 Tim-3 and Gal-9 are novel and promising predictors for post-transplant infection. • Relative decrease rate of Gal-9 shows potential to identify the risk of post-transplant infection. • Pre-transplant serum Tim-3 and Gal-9 shows no predictive power to post-transplant infection. [ABSTRACT FROM AUTHOR]