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Background: The incidence of early-onset colorectal cancer (EOCRC; diagnosed <50 years of age) is rising globally; however, the causes underlying this trend are largely unknown. CRC has strong genetic and environmental determinants, yet common genetic variants and causal modifiable risk factors underlying EOCRC are unknown. We conducted the first EOCRC-specific genome-wide association study (GWAS) and Mendelian randomization (MR) analyses to explore germline genetic and causal modifiable risk factors associated with EOCRC., Patients and Methods: We conducted a GWAS meta-analysis of 6176 EOCRC cases and 65 829 controls from the Genetics and Epidemiology of Colorectal Cancer Consortium (GECCO), the Colorectal Transdisciplinary Study (CORECT), the Colon Cancer Family Registry (CCFR), and the UK Biobank. We then used the EOCRC GWAS to investigate 28 modifiable risk factors using two-sample MR., Results: We found two novel risk loci for EOCRC at 1p34.1 and 4p15.33, which were not previously associated with CRC risk. We identified a deleterious coding variant (rs36053993, G396D) at polyposis-associated DNA repair gene MUTYH (odds ratio 1.80, 95% confidence interval 1.47-2.22) but show that most of the common genetic susceptibility was from noncoding signals enriched in epigenetic markers present in gastrointestinal tract cells. We identified new EOCRC-susceptibility genes, and in addition to pathways such as transforming growth factor (TGF) β, suppressor of Mothers Against Decapentaplegic (SMAD), bone morphogenetic protein (BMP) and phosphatidylinositol kinase (PI3K) signaling, our study highlights a role for insulin signaling and immune/infection-related pathways in EOCRC. In our MR analyses, we found novel evidence of probable causal associations for higher levels of body size and metabolic factors-such as body fat percentage, waist circumference, waist-to-hip ratio, basal metabolic rate, and fasting insulin-higher alcohol drinking, and lower education attainment with increased EOCRC risk., Conclusions: Our novel findings indicate inherited susceptibility to EOCRC and suggest modifiable lifestyle and metabolic targets that could also be used to risk-stratify individuals for personalized screening strategies or other interventions., (Copyright © 2024. Published by Elsevier Ltd.)

Objectives: National-level data on the incidence of retinopathy of prematurity (ROP) in different regions of China is insufficient. This study aimed to compare ROP incidences and care practices in different regions of China and their relationship with regional gross domestic product (GDP) per capita., Study Design: Retrospective cohort study., Methods: All infants born at <32 weeks gestational age (GA) and admitted to 70 neonatal intensive care units (NICUs) from January 1, 2019, to December 31, 2020, were enrolled. Hospitals were categorised into three regional groups according to geographical locations and GDP per capita from high to low: Eastern, Central, and Western China. The incidence of death or ROP, and care practices were compared among the groups., Results: A total of 18,579 infants were enrolled. Median GA was 29.9 (interquartile range 28.4-31.0) weeks and birth weight was 1318.1 (317.2) g. The percentage of GA <28 weeks, complete administration of antenatal steroids, and weight gain velocity during NICU stay were highest in Eastern China and lowest in Western China (all P < 0.01). In Eastern, Central, and Western China, the rates of death or any stage of ROP were 33.3%, 38.5%, and 39.2%, respectively (P < 0.01)., Conclusions: There were considerable regional disparities in ROP incidence in preterm infants with GA <32 weeks in China. The incidence of death or ROP ranged from high to low in Western, Central, and Eastern China., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Purpose: Currently there is a lack of effective treatment strategies for malignant superior vena cava syndrome (SVCS). We aim to investigate the therapeutic effect of intra-arterial chemotherapy (IAC) combined with the Single Needle Cone Puncture method for the 125 I brachytherapy (SNCP- 125 I) in treating SVCS caused by stage III/IV Small Cell Lung Cancer (SCLC)., Materials and Methods: Sixty-two patients with SCLC who developed SVCS from January 2014 to October 2020 were investigated in this study. Out of these 62 patients, 32 underwent IAC combined with SNCP- 125 I (Group A) and 30 patients received IAC treatment only (Group B). Clinical symptom remission, response rate, disease control rate, and overall survival of these two groups of patients were analyzed and compared., Results: The remission rate of symptoms including dyspnea, edema, dysphagia, pectoralgia, and cough of malignant SVCS in Group A was significantly higher than that in Group B (70.5 and 50.53%, P=0.0004, respectively). The disease control rates (DCR, PR+CR+SD) of Group A and B were 87.5 and 66.7%, respectively (P=0.049). Response rates (RR, PR+CR) of Group A and Group B were 71.9 and 40% (P=0.011). The median overall survival (OS) of Group A was significantly longer than that in Group B which was 18 months compared to 11.75 months (P=0.0360)., Conclusions: IAC treatment effectively treated malignant SVCS in advanced SCLC patients. IAC combined with SNCP- 125 I in the treatment of malignant SVCS caused by SCLC showed improved clinical outcomes including symptom remission and local tumor control rates than IAC treatment only in treating SCLC-induced malignant SVCS., (Copyright © 2023 Société française de radiothérapie oncologique (SFRO). Published by Elsevier Masson SAS. All rights reserved.)

Background and Aims: Diabetic retinopathy (DR) is one of the most common microvascular complications of diabetes mellitus (DM), and could increase the risks of adverse cardiovascular events among DM patients. Since heart failure with preserved ejection fraction (HFpEF) and DM often coexist, our present study aimed to explore the associations of DR with adverse outcomes in HFpEF patients., Methods and Results: We conducted this study in a large, international population suffering from HFpEF (n = 3442) based on the Treatment of Preserved Cardiac Function Heart Failure With an Aldosterone Antagonist (TOPCAT) trial. The associations of baseline DR with clinical outcomes were expressed as adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) using the Cox proportional hazard regression models. The crude incidence rates of all the outcomes studied were significantly increased when DM patients with or without DR compared to those without DM (all P < 0.05), whereas there were no differences between DM patients without DR versus those with DR (all P > 0.05). In the multivariate cox regression analysis, DR was not significantly associated with increased risks of the primary composite outcome (HR, 1.178 [95% CI, 0.870-1.596]) and secondary outcomes including all-cause death, cardiovascular death, all-cause hospitalization, hospitalization for HF, myocardial infarction, and stroke (all P > 0.05)., Conclusions: Our results of current study suggested that DM but not DR could be regarded as an independent risk factor for the prognosis of HFpEF., Clinical Trial Registration: URL: https://clinicaltrials.gov. Unique identifier: NCT00094302., Competing Interests: Declaration of competing interest All authors declare that they have no conflict of interest., (Copyright © 2022 The Italian Diabetes Society, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

Background: In the randomized phase III KEYNOTE-181 study, pembrolizumab prolonged overall survival (OS) compared with chemotherapy as second-line therapy in patients with advanced esophageal cancer and programmed death-ligand 1 (PD-L1) combined positive score (CPS) ≥10. We report a post hoc subgroup analysis of patients with esophageal squamous cell carcinoma (ESCC) enrolled in KEYNOTE-181 in Asia, including patients from the KEYNOTE-181 China extension study., Patients and Methods: Three hundred and forty Asian patients with advanced/metastatic ESCC were enrolled in KEYNOTE-181, including the China cohort. Patients were randomly assigned 1 : 1 to receive pembrolizumab 200 mg every 3 weeks for ≤2 years or investigator's choice of paclitaxel, docetaxel, or irinotecan. OS, progression-free survival, response, and safety were analyzed without formal comparisons. OS was evaluated based on PD-L1 CPS expression level., Results: In Asian patients with ESCC, median OS was 10.0 months with pembrolizumab and 6.5 months with chemotherapy [hazard ratio (HR), 0.63; 95% CI 0.50-0.80; nominal P < 0.0001]. Median progression-free survival was 2.3 months with pembrolizumab and 3.1 months with chemotherapy (HR, 0.79; 95% CI 0.63-0.99; nominal P = 0.020). Objective response rate was 17.1% with pembrolizumab and 7.1% with chemotherapy; median duration of response was 10.5 months and 7.7 months, respectively. In patients with PD-L1 CPS <1 tumors (pembrolizumab versus chemotherapy), the HR was 0.99 (95% CI 0.56-1.72); the HR (95% CI) for death was better for patients with PD-L1 CPS cut-offs >1 [CPS ≥1, 0.57 (0.44-0.75); CPS ≥5, 0.56 (0.41-0.76); CPS ≥10, 0.53 (0.37-0.75)]. Treatment-related adverse events were reported in 71.8% of patients in the pembrolizumab group and 89.8% in the chemotherapy group; grade 3-5 events were reported in 20.0% and 44.6%, respectively., Conclusions: Pembrolizumab monotherapy demonstrated promising efficacy in Asian patients with ESCC, with fewer treatment-related adverse events than chemotherapy. PD-L1 CPS ≥1 is an appropriate cut-off and a predictive marker of pembrolizumab efficacy in Asian patients with ESCC., Competing Interests: Disclosure YC, SQ, SL, ZL, YC, YF, YS, XY, XY, WL, TL, XL, JZ, YB, CB, KG, H-MP, LB, J-WY, JC have declared no conflicts of interest. C-HH reports honoraria and fees for consulting and lecture from Bristol Myers Squibb, Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA, Ono Pharmaceutical, and Roche; fees for consulting from Merck Serono; and grants from AstraZeneca and BeiGene. S-BK reports grants (to institution) from DongKook Pharm Co., Novartis, and Sanofi Genzyme; and personal fees (advisory) for Dae Hwa Pharma Co. Ltd, Daiichi-Sankyo, ISU Abxis. TK reports grants and personal fees from Astellas Amgen BioPharma, Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA, and Ono Pharmaceutical; grants from Shionogi and Taiho Pharmaceutical; and personal fees from Astellas Pharma, Bristol Myers Squibb, and Oncolys BioPharma. S-HL reports personal fees from AstraZeneca, Bristol Myers Squibb, Merck Sharp & Dohme Corp., and Roche; and grants from Merck Sharp & Dohme Corp., KM reports research funding for Amgen, Daiichi-Sankyo, Merck Serono, Merck Sharp & Dohme Corp., Ono, Parexel International, Pfizer, Sanofi, Solasia Pharma, and Taiho; personal fees (consulting fee and/or honorarium) for Amgen, AstraZeneca, Bayer, Bristol Myers Squibb, Chugai, Eli Lilly, Ono, Sanofi, Taiho, and Takeda. TD reports grants from AbbVie, Boehringer Ingelheim, Bristol Myers Squibb, Daiichi-Sankyo, Eisai, Janssen, Lilly, Merck Serono, Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA, Novartis, Pfizer, Quintile (IQVIA), Sumitomo Dainippon, and Taiho; and personal fees from AbbVie, Amgen, Astellas Pharma, Bayer, Boehringer Ingelheim, Bristol Myers Squibb, Chugai Pharma, Daiichi Sankyo, Janssen, Merck Sharp & Dohme Corp., Novartis, Oncolys BioPharma, Ono Pharmaceutical, Otsuka Pharma, Rakuten Medical, Sumitomo Dainippon, Taiho, and Takeda. YC is an employee of MSD China, Shanghai, China. WL is an employee of MSD China, Shanghai, China., (Copyright © 2021 Merck Sharp & Dohme Corp. , a subsidiary of Merck & Co., Inc., Kenilworth, NJ USA, The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Although colorectal cancer (CRC) incidence in the USA is declining overall, its incidence is increasing among those younger than 50 years of age. The reasons underlying the increasing trend are largely unknown, although behavioral changes, such as unhealthy diet and lifestyle factors, may be partially responsible., Design: A prospective cohort study included 94 217 women aged 26-45 years at baseline. Validated anthropometric measures and lifestyle information were self-reported biennially. Exposures were four recommendation-based dietary indices-the prime diet quality score and three plant-based dietary indices; and two mechanism-based indices-the empirical dietary and lifestyle index for hyperinsulinemia (EDIH and ELIH). We calculated hazard ratios (HRs) and 95% confidence intervals (CIs) for overall CRC and for early-onset (before age 50) and after age 50 CRC separately., Results: We documented 332 cases of CRC during 24 years of follow-up (2 113 655 person-years), with an average age of 52 ± 7 years at diagnosis. Hyperinsulinemic dietary and lifestyle patterns were associated with a higher risk of CRC. Multivariable-adjusted HRs (95% CIs) comparing participants in the highest versus lowest quartile were: 1.67 for EDIH (95% CI: 1.15-2.44, P-trend = 0.01) and 1.51 for ELIH (95% CI: 1.10-2.08, P-trend = 0.01). Moreover, per 75% increment in rank, ELIH appeared to be a stronger risk factor for early-onset CRC (HR = 1.86, 95% CI: 1.12-3.07) than after age 50 CRC (HR = 1.20, 95% CI: 0.83-1.73, P-heterogeneity = 0.16). The four recommendation-based indices were not significantly associated with overall, early-onset, or after age 50 CRC risk (per 75% increment in rank, HRs ranged from 0.75 to 1.28)., Conclusion: Dietary and lifestyle patterns contributing to hyperinsulinemia were associated with greater CRC risk in younger women. Moreover, the hyperinsulinemic lifestyle showed a suggestively stronger positive association with early-onset CRC risk, compared with after age 50 CRC. Our findings suggest that dietary and lifestyle interventions to reduce insulinemic potential may be effective for CRC prevention among younger women., Competing Interests: Disclosure JAM has received institutional research funding from Boston Biomedical, has served as an advisor/consultant to Ignyta and COTA Healthcare, and served on a grant review panel for the National Comprehensive Cancer Network funded by Taiho Pharmaceutical. All other authors have declared no conflicts of interest. Role of the funder/sponsor The funders had no role in the design and conduct of the study; collection, management, analysis, and interpretation of the data; preparation, review, or approval of the manuscript; and decision to submit the manuscript for publication., (Copyright © 2021 European Society for Medical Oncology. All rights reserved.)

Objectives: Sleep disorder is a common problem in children that can jeopardize their health and well-being. With the popularity of electronic devices such as portable tablets and smartphones in the 21st century, children are spending much more time on screen, but the impact of such change on children's sleep disorder has been less investigated so far. This study aims to examine the dose-response association between time spent on different electronic devices and children's sleep disorder., Study Design: The design of this study is a cross-sectional study., Methods: We randomly selected 2278 children aged 3-6 years from 15 kindergartens in Tongling, China. The potentially non-linear association between screen-viewing time (i.e. television [TV], computer, iPad, Phone) and the risk of sleep disorder was examined using a logistic generalized additive model., Results: We observed a J-shaped association between TV viewing time and the risk of sleep disorder, with a threshold of 1 h/day. For each 1 h/day increment in TV viewing time over the threshold, the risk of sleep disorder increased by 12.35% (95% confidence interval: 1.87-23.92%). This association seemed to be greater for girls than boys and for TV viewing at weekend than on weekdays, but the difference was not statistically significant (P-value>0.05). We did not find adequate evidence of an adverse effect of more time spent on computer, iPad and Phone., Conclusions: This study suggests a positive but non-linear relationship between time spent on watching TV and sleep disorder in Chinese preschool children. Setting the TV viewing time limit less than 1 h/day may help reduce the risk of developing sleep disorder. Further investigation is also needed to examine and compare the effects of heavy use of other electronic devices on sleep disorder., (Copyright © 2020 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

With intensive selection for meat production in broilers, excessive fat accumulation is also accompanied and causes economic concerns. Folic acid was reported to be involved in lipid metabolism. The present study was conducted to investigate the role of folic acid in reducing abdominal fat deposition. A total of 105 one-day-old healthy Arbor Acres broilers were randomly distributed into 3 treatments, including the control (Con), saline-perfusion group (NS), and folic acid perfusion group (FA). The growth performance, biochemical characteristics in serum, and lipid metabolism in the liver and abdominal fat tissues were evaluated. Results have shown that folic acid significantly reduced abdominal fat percentage (P < 0.05) and had no effects on BW, ADFI, ADG, and FCR (P > 0.05). Serum triglycerides (TG), total cholesterol (TC), and alanine aminotransferase (ALT) levels were lower in FA group but albumin concentration was higher (P < 0.05). Hepatic ACC, SCD, ELOVL6, PI3K, LDLR, HMGCR, and ABCA1 mRNA abundance were all down-regulated in FA group (P < 0.05) when compared with the Con and NS groups, while CPT1 and PPARα were not affected. In addition, MTTP mRNA abundance was higher in the liver of birds subjected to folic acid (P < 0.05). There was no difference about TG deposition in the liver among all groups based on hematoxylin-eosin (HE) and Oil Red O staining. On the other hand, ELOVL6, PPARγ, IGF1, and TGFβ2 expression were notably decreased in the abdominal fat in FA group (P < 0.05). In conclusion, our data demonstrated that folic acid has reduced abdominal fat percentage by decreasing hepatic lipogenesis and suppressing adipocytes proliferation and differentiation. And the inhibiting effect of adipocytes might be mediated by IGF1 and TGFβ2 down-regulation., (© 2019 Poultry Science Association Inc.)

Hypoxia-inducible factor 1α (HIF1α) activation is profoundly implicated in the initiation and progression of multiple malignant tumors. Prolyl 4-hydroxylase subunit alpha 1 (P4HA1) is the active catalytic component of prolyl 4-hydroxylase and has been reported to promote tumor progression in several cancers. In this study, we revealed that P4HA1 was highly expressed in pancreatic ductal adenocarcinoma (PDAC) and predicted a poor clinical outcome. Notably, elevated expression of P4HA1 in PDAC cells was HIF1α-dependent. Gene set enrichment analysis of The Cancer Genome Atlas (TCGA) cohort demonstrated a close link between P4HA1 expression and glycolysis and hypoxia gene signatures in PDAC. Knockdown of P4HA1 significantly suppressed the glycolytic activity of PDAC cells as revealed by reduced glucose utilization and lactate production. Consistently, there was a close correlation between P4HA1 and glycolysis genes. Moreover, we found that P4HA1 can enhance HIF1α stability, indicating a positive feedback loop between HIF1α and P4HA1 in PDAC. Genetic silencing of P4HA1 significantly inhibited the cell proliferation, chemoresistance, and stemness of PDAC cells. Collectively, our findings identify the P4HA1-HIF1α loop as a critical regulator in glycolysis and oncogenic activities of PDAC and provide a potential target for pancreatic cancer treatment., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The high mobility group A1 (HMGA1) protein, an architectural transcription factor, is profoundly implicated in the pathogenesis and progression of multiple malignant tumors. Reprogrammed energy metabolism is a hallmark of diverse types of cancer cells. However, little is known about the regulatory role of HMGA1 in aerobic glycolysis. In this study, we found that HMGA1 was highly expressed in many types of human cancers including gastric cancer and predicted a poor prognosis. However, high HMGA1 expression was not correlated with TNM stages. Gene set enrichment analysis result suggested a link between HMGA1 expression and glycolytic phenotype in gastric cancer. Genetic silencing of HMGA1 significantly inhibited gastric cancer glycolytic activity as revealed by reduced glucose uptake, lactate release, and extracellular acidification ratio. In addition, cell proliferation and invasive capacity of gastric cancer cells were also suppressed by HMGA1 knockdown. Mechanistically, the key glycolysis regulator c-Myc was identified as a downstream target of HMGA1. In gastric cancer patients, HMGA1 and c-Myc expression were closely associated with the glycolysis gene signature. Taken together, our findings identify a novel function of HMGA1 in regulating aerobic glycolysis in gastric cancer., (Copyright © 2019. Published by Elsevier Inc.)

A virulent infectious bronchitis virus (IBV), designated as CK/CH/GD/QY16 (referred as QY16), was isolated from a diseased chicken farm in Guangdong province, China, in 2016. The complete genome of the strain was sequenced and analyzed. The results show that the genome of QY16 consists of 27,670 nucleotides, excluding poly (A) tail, and that its genome organization is 5' UTR-1a-1b-S-3a-3b-E-M-4b-4c-5a-5b-N-6b-3' UTR-poly (A) tail. Sequence comparison among QY16 and other IBV strains was conducted and its results demonstrate that the S1 gene of QY16 has the highest nucleotide sequence identity with that of 4/91, and the other part of its genome is highly similar to that of YX10. The results of the phylogenic analysis show that the entire genome of QY16 and most of the QY16 genes are located in the same cluster as those of YX10, except for the S1 gene which is located in the same cluster with that of 4/91. It has been further confirmed by the RDP and SimPlot analysis that QY16 is a recombinant strain deriving from YX10 (as the major parental sequence) and 4/91 (as the minor parental sequence), and that the recombination occurs in a region which includes the 3'-terminal 1b sequence (85 nt) and the 5'-terminal S1 protein gene sequence (1,466 nt). The results of the vaccination-challenge test suggest that QY16 is a nephropathogenic strain of IBV and that the vaccine strains-H120 and 4/91-cannot provide effective protection against it. These results indicate that the continuing evolution of IBV strains by genetic drift and genetic recombination may lead to IBV outbreaks even among the vaccinated chickens in China.

Background & Aims: A growing body of evidence suggests that beta-glucan derived from oats or barley can reduce cardiovascular disease risk through reductions in serum lipids. However, the effects of beta-glucan on lipid changes in hypercholesterolemic patient groups are inconsistent. The objective of this study was to identify and quantify the effect of beta-glucan, a marker of water-soluble fiber, on various lipid parameters and glucose level in hypercholesterolemic subjects., Methods and Results: We performed a comprehensive literature search to identify the relevant randomized controlled trials (RCTs) that investigated the effects of beta-glucan consumption in hypercholesterolemic subjects. Mean differences (MDs) and 95% confidence intervals (CIs) were calculated for net changes in lipid concentrations by using fixed-effects or random-effects models according to heterogeneity. Publication bias, sensitivity analysis and subgroup analyses were also performed. Seventeen eligible RCTs with 916 subjects were included in the meta-analysis. The pooled result showed that beta-glucan consumption in hypercholesterolemic population significantly lowered the total cholesterol (TC) (MD, -0.26 mmol/L; 95% CI, -0.33 to -0.18; P < 0.00001) and low-density lipoprotein (LDL)-cholesterol concentration (MD, -0.21 mmol/L; 95% CI, -0.27 to -0.14; P < 0.00001). However, there were no significant differences in high-density lipoprotein (HDL)-cholesterol, triglycerides (TG) and glucose. No adverse effects were reported among the eligible trials., Conclusion: Our meta-analysis showed that beta-glucan consumption significantly decreased TC and LDL-cholesterol concentrations but did not affect TG, HDL-cholesterol, and glucose concentrations in hypercholesterolemic subjects., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In March 2013, an influenza outbreak caused by the novel avian-origin H7N9 influenza A virus emerged in eastern China and had caused 43 fatalities by 31 July 2013, although the basis for disease pathogenesis still remains unclear. To assess the immunological and viral factors associated with disease severity, viral RNA and inflammatory cytokines were quantified for 18 H7N9 patients in Shanghai, China. Detailed clinical information was collected and clinical laboratory investigations were performed for all patients. H7N9 infection is characterized by high pharyngeal virus load and frequent detection of viral RNA in blood. High pharyngeal virus load persisted through the first 10 days of antiviral therapy in fatal cases. Genetic characterization of the H7N9 virus revealed an Arg292Lys mutation in the neuraminidase gene associated with oseltamivir-resistance. Pronounced lymphopenia and high chemokine and cytokine levels were observed in H7N9-infected patients, particularly in those where disease was fatal. Serum levels of interleukin-6 (IL-6), IL-8 and macrophage inflammatory protein-1β in our subjects also correlated positively with pharyngeal virus load. Lymphocyte counts <0.5 × 10(9)  cells/L, and serum IL-6 >97 pg/mL, IL-8 >40 pg/mL and C-reactive protein >90 mg/L were identified as being connected with adverse clinical outcome through univariate logistic analysis. Significant survival differences were also observed between patients with serum C-reactive protein <90 mg/L or creatinine <90 μmol/L and those with higher levels. Our data demonstrated that high viral load, and the resulting intense inflammatory responses, played an important role in H7N9 pathogenesis. Though immunomodulatory treatment has potential benefits, the focus of clinical management should be on preventing the intense cytokine response by early diagnosis and effective antiviral treatment., (© 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.)

In China, KSHV seroprevalence varies considerably among different regions and ethnicities. But in Xinjiang province, located in the northwestern China, there is a very high seroprevalence of KSHV in adults of Kazak and Ughur ethnicities. However, KSHV prevalence in children and the risk factors associated with the acquisition of infection are currently not known. The aim of this study was to investigate the prevalence of KSHV infection and identify associated socioeconomic or behavioural risk factors and the humoral immune response among children in this population. This is a cross-sectional study (N = 178) to screen children and their caregivers from Xinjiang for total KSHV antibodies, KSHV neutralizing antibodies and HIV infection. Structured questionnaires were utilized to investigate risk factors associated with KSHV prevalence. KSHV seroprevalence in children and caregivers in Xinjiang was 48.3% and 64.7%, respectively. Neutralizing antibody was detected in most seropositive caregivers (93.8%) but was detected in only 5.8% of the infected children. A significant association was observed between child KSHV seroprevalence and sharing of food among family members. These results suggest that similar to other endemic areas in Africa, KSHV infection in the minority populations of Xinjiang is likely to be occurring during early childhood, probably via horizontal transmission through saliva, and results in high seroprevalence in the adult population., (© 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.)

Objectives: This ex vivo study compared the physico-chemical structural differences between primary carious teeth biannually treated with silver diamine fluoride (SDF) and carious teeth without such treatment., Method: Twelve carious primary upper-central incisors were collected from 6-year-old children. Six teeth had arrested caries after 24-month biannual SDF applications and 6 had active caries when there was no topical fluoride treatment. The mineral density, elemental contents, surface morphology, and crystal characteristics were assessed by micro-computed tomography (micro-CT), energy-dispersive X-ray spectrometry (EDX), scanning electron microscopy (SEM), and transmission electron microscopy (TEM)., Results: Micro-CT examination revealed a superficial opaque band approximately 150μm on the arrested cavitated dentinal lesion. This band was limited in the active carious lesion. EDX examination detected a higher intensity of calcium and phosphate of 150μm in the surface zone than in the inner zone, but this zone was restricted in the active cavitated dentinal lesion. SEM examination indicated that the collagens were protected from being exposed in the arrested cavitated dentinal lesion, but were exposed in the active cavitated dentinal lesion. TEM examination suggested that remineralised hydroxyapatites were well aligned in the arrested cavitated dentinal lesion, while those in the active cavitated dentinal lesion indicated a random apatite arrangement., Conclusions: A highly remineralised zone rich in calcium and phosphate was found on the arrested cavitated dentinal lesion of primary teeth with an SDF application. The collagens were protected from being exposed in the arrested cavitated dentinal lesion., Clinical Significance: Clinical SDF application positively influences dentine remineralisation., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Aim: The expression of cysteine cathepsins in human carious dentine suggests that this enzyme contributes to the collagen degradation in caries progress. This study investigated whether silver diamine fluoride (SDF) inhibited the activity of cysteine cathepsins., Methods: Three commercial SDF solutions with concentrations at 38%, 30% and 12% were studied. Two fluoride solutions with the same fluoride ion (F(-)) concentrations as the 38% and 12% SDF solutions, and 2 silver solutions with the same silver ion (Ag(+)) concentrations as the 38% and 12% SDF solutions were prepared. Five samples of each experimental solution were used to study their inhibitory effect on two cathepsins (B and K) using cathepsin assay kits. Positive control contained assay buffer and cathepsins dilution was used to calculate the percentage inhibition (difference between the mean readings of the test solution and control solution divided by that of the control group)., Results: The percentage inhibition of 38%, 30% and 12% SDF on cathepsin B were 92.0%, 91.5% and 90.3%, respectively (p<0.001); on cathepsin K were 80.6%, 78.5% and 77.9%, respectively (p<0.001). Ag(+) exhibited the inhibitory effect against both cathepsin B and K with or without the presence of F(-) (p<0.01). The solutions containing Ag(+) have significantly higher inhibitory effect than the solutions containing F(-) only (p<0.01)., Conclusion: According to this study, SDF solution at all 3 tested concentrations significantly inhibited the activity of cathepsin B and K., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Introduction: The optimal treatment of in-stent restenosis (ISR) is not determined. We describe the efficacy of carotid endarterectomy (CEA) with stent removal in the management of symptomatic ISR., Methods: A 72-year-old man presented with recurrent right limb weakness 8 months after carotid artery stenting. Digital substraction angiography showed severe ISR (almost 99%). CEA with stent removal was performed., Results: A total of 41 cases was reviewed: periarterial inflammation was shown in 10.3% of patients; 92.9% had clear intima-media plane; and 85.4% had a good outcome. All patients remained stable at follow-up., Discussion: CEA with stent removal appears to be a safe, feasible, effective, and durable therapeutic option., (Copyright © 2013 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.)

Objective: To investigate the inhibitory effects of 38% silver diamine fluoride (SDF) on demineralised dentine., Methods: Human dentine blocks were demineralised and allocated to four groups: SF, F, S and W. The blocks in group SF received a topical application of 38% SDF solution (253,900ppm Ag, 44,800ppm F), group F received a 10% sodium fluoride solution (44,800ppm F), group S received a 42% silver nitrate solution (253,900ppm Ag) and group W received deionised water (control). They were subjected to pH cycling using demineralisation solution (pH 5) and remineralisation solution (pH 7) for 8 days. The surface morphology, crystal characteristics, lesion depth and collagen matrix degradation of the specimens were investigated by scanning electron microscopy (SEM), X-ray diffraction (XRD), micro-CT testing and spectrophotometry with a hydroxyproline assay., Results: The surface morphology under SEM showed evident demineralisation with exposed collagen in groups S and W, but not in group SF. Clusters of granular spherical grains were observed in the cross-sections of specimens in groups SF and F. XRD revealed precipitates of silver chloride in groups SF and S. The mean lesion depths (±SD) of groups SF, F, S and W were 182 ± 32μm, 204 ± 26μm, 259 ± 42μm and 265 ± 40μm, respectively (SDF, F

Atom probe is a technique that is highly suited to the study of nanocrystalline materials. It can provide accurate atomic-scale information about the composition of grain boundaries in three dimensions. In this paper we have analysed the microstructure of a nanocrystalline super-duplex stainless steel prepared by high pressure torsion (HPT). Not all of the grain boundaries in this alloy display obvious segregation, making visualisation of the microstructure challenging. In addition, the grain boundaries present in the atom probe data acquired from this alloy have complex shapes that are curved at the scale of the dataset and the interfacial excess varies considerably over the boundaries, making the accurate characterisation of the distribution of solute challenging using existing analysis techniques. In this paper we present two new data treatment methods that allow the visualisation of boundaries with little or no segregation, the delineation of boundaries for further analysis and the quantitative analysis of Gibbsian interfacial excess at boundaries, including the capability of excess mapping., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Implantable microelectrode arrays (MEA) hold enormous hope for individuals with sensory or motor deficits. However, long-term function of MEA remains a critical hurdle. The objective of this study was to synthesize an antioxidant prodrug that can be delivered to the neural tissue around the implant and present a pharmacological depot to combat the injurious oxidative stress around the MEA. In this report, monomers of triethylene glycol methyl acrylate and α-tocopheryl acrylate, a synthetic derivative of the antioxidant α-tocopherol (vitamin E, Ve), were copolymerized to obtain poly(triethylene glycol methyl acrylate-co-α-tocopheryl acrylate) (PVT) with different compositions. In contrast to the poor water solubility of Ve, solubility of the PVT prodrug in water can reach as high as 3.1 mg ml(-1) (equivalent to 500 μM Ve) by tuning the copolymer composition. To demonstrate the applicability of the prodrug for MEA implants, PVT was successfully deposited on silicon substrates with poly(acrylic acid) (PAA) or tannic acid (TA) using the layer-by-layer technique mediated by hydrogen bonding. Ellipsometry and quartz crystal microbalance data showed that the multilayers of PAA/PVT were destructible at physiological pH. In contrast, multilayers of TA/PVT were stable. The PVT prodrug was non-cytotoxic toward A172 human astrocytes. Furthermore, PVT was able to protect astrocytes against oxidative stress exerted by H(2)O(2) in vitro. Using a free radical scavenging assay, the protection mechanism was attributed to the hydrolysis of the labile ester linkage and release of the active Ve., (Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The objective of this study was to identify alternative transcripts and single nucleotide polymorphisms (SNPs) in the 3'-untranslated region (3' UTR) of bovine malic enzyme (ME1) gene and to evaluate the extent to which polymorphisms were associated with meat quality and carcass traits in Chinese red cattle. Two transcripts, long transcript and short transcript that differ in the length of the 3' UTR were cloned. A single nucleotide polymorphism was detected in 3' UTR and a restriction site for endonuclease ME1-Dra I was also found. The result revealed that the ME1-Dra I genotypes had a significant effect on cooking loss, pH measured 24h post-mortem (pH(24h)) and eye muscle area (P < 0.05). In conclusion, the SNPs may be used as DNA markers to select for meat quality and carcass traits in Chinese red cattle., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

We investigated the effect of homogentisate 1, 2 dioxygenase (HGD) gene on meat quality and carcass traits in 287 Chinese red cattle. The PCR-SSCP method was used to identify polymorphism of the HGD gene in the exon 1 and intron 1. Two polymorphisms were detected in intron 1 and two restriction sites for endonuclease HGD-BstXI and HGD-HaeIII have also been found. The HGD-BstXI genotypes showed significant effects on cooking loss, drip loss, net meat weight, carcass weight, and eye muscle area (P<0.05). The HGD-HaeIII genotypes significant affected cooking loss, muscle fibre diameter, shear force, drip loss, and carcass yield ratio (P<0.05). Moreover, we found significant effects of diplotypes on cooking loss, muscle fibre diameter, shear force, drip loss, net meat weight, carcass weight, and eye muscle area (P<0.05)., (Copyright 2010 Elsevier Ltd. All rights reserved.)

This prospective study aims to evaluate the impact of the excimer laser technology as the first-line endovascular treatment of critical limb ischaemia (CLI) in diabetic patients. The protocol allowed the use of laser ablation of obstructive lesions when conventional endoluminal guidewire crossing of the plaque was unsuccessful. We extrapolate the data of consecutive patients treated, who completed at least 12 months of follow-up, extending the observation to a 26-month time frame. During this period, 67 diabetic patients with CLI were brought to the Cath Lab for 'operative angioplasty' and to be treated with endovascular techniques. Of the 67 cases, laser was used on 35 patients to treat 51 lesions. All patients had type C or D occlusive lesions, according to the TACS II classification, showing a single type D plaque or multiple tandem C/D occlusive plaques ranging from 4 to 23 cm in length. The immediate clinical success, defined as restored direct arterial flow to the foot, was 88.2%. The lesions were successfully crossed by laser in 45 out of 51 attempts. Stents were required in 25% of the patients with 21% lesions. Patency rates were assessed using the Kaplan-Meier survival curves. The patency rates of the successfully treated lesions (freedom from target lesion revascularisation) were 96.6% at 12 months and 82.7% at 24 months. Limb-salvage rate at 12 and 24 months were 100% and 94%, respectively. Our study showed that the excimer laser-assisted angioplasty, when feasible, is effective in granting event-free survival in CLI patients with diabetes, and that endoluminal-driven atherectomy allows long-term success in reducing the need of stents in the lower limb arteries., (Copyright (c) 2009 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.)

Background: Data on the impact of gastroesophageal reflux disease (GERD) on health-related quality of life (HRQL) in Asian countries are scarce., Aim: This study evaluated the impact of GERD on HRQL in Shanghai, China., Subjects: One thousand two hundred adult inhabitants of Shanghai, selected using randomized cluster sampling., Methods: Participants completed Mandarin versions of the Reflux Disease Questionnaire (RDQ), GERD impact scale, quality of life in reflux and dyspepsia (QOLRAD) questionnaire and short-form-36 (SF-36). GERD was defined as heartburn and/or regurgitation of any frequency during the 1-week recall period of the RDQ. A clinically meaningful impairment of HRQL was defined as a statistically significant decrease of >or=0.5 points in a QOLRAD dimension or >or=5 points in an SF-36 dimension., Results: Overall, 1034 subjects completed the survey (86.2% response rate); 919 responses were suitable for analysis. The prevalence of GERD was 6.2%. GERD was associated with meaningfully impaired HRQL in the QOLRAD dimensions of vitality, eating/drinking and emotional well-being, but not sleep or physical/social functioning, and in all SF-36 dimensions except social functioning. Respondents with GERD experienced eating and drinking problems (47%), sleep impairment (32%) and reduced work productivity (32%)., Conclusion: GERD has a clinically meaningful impact on HRQL in Shanghai, China.

Staphylococcus aureus isolates (n = 30) from the environment or from surgical patients in a hospital in Guangzhou, China were investigated for the presence of class 1 integrons. The class 1 integrase (intI1) gene and its 3' conserved segment were detected by PCR. IntI1-positive isolates were further analysed for the presence of resistance gene cassettes using specific primers, intI1-K and In-B. All isolates were also subjected to multilocus sequence typing (MLST) and random amplified polymorphic DNA (RAPD)-PCR analysis. Sixteen (53%) clinical and environmental isolates were positive for the class 1 integrase gene and were also found to possess the aadA2 gene. The 30 isolates were classified into six distinct genotypes by RAPD-PCR analysis: type A (n = 2); type B (n = 2); type C (n = 3); type D (n = 7); type E (n = 8); and type F (n = 8). All isolates belonged to the same sequence type (ST239) by MLST. These results indicated transmission of S. aureus between the environment and patients, as well as the probability of nosocomial infection.

We investigated changes of tensile strength in tendon repair according to tension direction. Thirty-six fresh-frozen digital flexor tendons were divided into 4 groups with 9 tendons each. The tendons were repaired by the modified Kessler method. Sutured tendons were pulled against pulleys at angles of 0 degrees, 30 degrees, 60 degrees, and 90 degrees to the direction of the pull of the testing machine in the 4 groups, respectively. The repaired tendons were tested in a tensile machine to determine 2-mm gap formation force and ultimate strength of the tendons. The 2-mm gap formation force and ultimate strength in the tendons pulled at 0 degrees were statistically higher than those in the tendons pulled at 30 degrees, 60 degrees, and 90 degrees. The 2-mm gap formation force of the tendons pulled at 30 degrees, 60 degrees, and 90 degrees was 86% +/- 10%, 73% +/- 9%, and 64% +/- 8% of that at 0 degrees, respectively. Ultimate strength of tendons pulled at 30 degrees, 60 degrees, and 90 degrees was 89% +/- 9%, 82% +/- 11%, and 76% +/- 8% of that at 0 degrees, respectively. Values of the 2-mm gap formation force and ultimate strength were statistically the lowest in the group with a pulling angle of 90 degrees. There was no statistically significant difference in repair strength between tendons tested at 0 degrees and those in the model without pulleys. The strength of tendon repair changed considerably according to direction of tension added to the tendons. The gap formation force and ultimate strength decreased as angles of tension increased. The results imply that a repaired tendon will be weakened as the finger is increasingly flexed. The decrease in repair strength should therefore be considered in planning a tendon suture to tolerate active finger flexion and a tendon motion protocol after primary tendon repair.

Ornithine decarboxylase (ODC) is involved in the biosynthesis of polyamines and hence has been found in almost all types of cells studied. Therefore it is frequently used as internal standard. We isolated a cDNA, XODC2, which is a paralogue to ubiquitous ODC and expressed in a spatial and temporal manner during the early embryogenesis of Xenopus laevis. Expression of XODC2was first detected at the animal pole at stage 9. During neurula stages the signals were found both in the extreme anterior and posterior part of the dorsal body axis. In tailbud stages the expression is further shifted to both the tail and head areas and gradually restricted to distinct tissues: forebrain, inner layer of epidermis of the head area, stomodeal-hypophyseal anlage, frontal gland, ear vesicle, branchial arches, the front tip of neural tube and proctodeum. In addition, signals were also found in the inner layer of epidermis underneath the cement gland during early tailbud stages while in later tailbud stages signals were detected at the apical zone of the cement gland. Comparative studies indeed could confirm that XODC1 in contrast to XODC2 is expressed ubiquitously throughout the whole embryos during early development of Xenopus laevis.

A number of endogenous inhibitors targeting the tumor vasculature have recently been identified using in vitro and in vivo antiangiogenesis models. While many of these angiogenesis inhibitors display a broad spectrum of biological actions on several systems in the body, several inhibitors including angiostatin, endostatin, and serpin antithrombin seem to act specifically on the proliferating endothelial cell compartment of the newly formed blood vessels. The discovery of these specific endothelial inhibitors not only increases our understanding of the functions of these molecules in the regulation of physiological and pathological angiogenesis, but may also provide an important therapeutic strategy for the treatment of cancer and other angiogenesis dependent diseases, including diabetic retinopathy and chronic inflammations. Systemic administration of these angiogenesis inhibitors in animals significantly suppresses the growth of a variety of tumors and their metastases. However, their production as functional recombinant proteins has been proven to be difficult. In addition, high dosages of these inhibitors are required to suppress tumor growth in animal studies. Other disadvantages of the antiangiogenic protein therapy include repeated injections, prolonged treatment, transmission of toxins and infectious particles, and high cost for manufacturing large amounts of protein molecules. Thus, alternative strategies need to be developed in order to improve the clinical settings of antiangiogenic therapy. Developments of these strategies are ongoing and they include identification of more potent inhibitors, antiangiogenic gene therapy, improvement of protein/compound half-lives in the circulation, increase of their concentrations at the disease location, and combinatorial therapies with approaches including chemotherapy, radiotherapy, and immunotherapy. Despite the above-mentioned disadvantages, a few inhibitors have entered into the early stages of clinical trials and they may bring new hopes for the treatment of cancer and other angiogenesis dependent diseases.

We have investigated the ability of dsRNA to inhibit gene functions in zebrafish using sequences targeted to the maternal gene pouII-1, the transgene GFP, and an intron of the zebrafish gene terra. We found that embryos injected with all of these dsRNAs at approximately 7.5 pg/embryo or higher had general growth arrest during gastrulation and displayed various nonspecific defects at 24 h postfertilization, although embryonic development was unaffected before the midblastula stage. Reducing dsRNA concentration could alleviate the global defects. Injection of GFP dsRNA (7.5-30 pg/embryo) did not inhibit GFP expression in transgenic fish, although abnormal embryos were induced. Co-injection of GFP mRNA with either GFP or non-GFP dsRNA caused reduction of GFP expression. Whole-mount in situ hybridization clearly showed that embryos injected with dsRNA degraded co-injected and endogenous mRNA without sequence specificity, indicating that dsRNA has a nonspecific effect at the posttranscriptional level. It appears that RNAi is not a viable technique for studying gene function in zebrafish embryos., (Copyright 2001 Academic Press.)

The oxidative stability of self-assembled monolayer (SAM) of thiols on silver was examined with matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS). A benzenesulfinate (BS) monolayer on silver was also prepared and investigated with MALDI-TOF-MS and surface-enhanced Raman scattering (SERS). The presence of sulfinate, sulfonate, and thiosulfonate fragment ions reveals that thiolate monolayers oxidize in the air. The relative abundance of C6H5SO2- and C6H5SO3- species in the mass spectra of the monolayer with different air-exposure times provides an estimation of the oxidation progress. In the vibrational spectrum, the large red shift of v(SOO-), combined with the lack of a shift of v(C-S) upon adsorption, indicates its bidentate O-coordination. The orientation of the confined molecules, based on the Raman surface selection rules, was derived from the preferential enhancement of the different functional groups. The benzene ring of BS was found almost normal to the metal surface as a result of the intermolecular forces; that geometry excludes the possibility of the pi-system of the benzene ring from participating as another binding site. The SER spectra of BS obtained via a selective etching process confirms these observations.

The aim of the present study was to observe the effect of repeated subcutaneous (sc) injections of low doses of ketamine for the treatment of acute inflammatory pain in a complete Freund's adjuvant-induced monoarthritic pain model in rats. The results show: (1) sc injection of ketamine at a dose of 2 mg x kg(-1) or 10 mg x kg(-1) produced significant analgesia (P<0.01) in arthritic rats starting from the 2nd week and 3rd week, respectively. (2) Repeated administration of ketamine produced a significant reduction of the circumference of the arthritic ankle (P<0.05 and P<0.01 with different doses). (3) The body weight of the rats was not affected by continuous administration of ketamine for 4 weeks. No abnormal locomotor behavior was observed (It was concerned but not systemically evaluated in this study). The results suggest that repeated sc injection of ketamine for 4 weeks significantly reduce inflammatory pain in monoarthritis without notable aversive side effects.

During primary HIV infection the viral load in plasma increases, reaches a peak, and then declines. Phillips has suggested that the decline is due to a limitation in the number of cells susceptible to HIV infection, while other authors have suggested that the decline in viremia is due to an immune response. Here we address this issue by developing models of primary HIV-1 infection, and by comparing predictions from these models with data from ten anti-retroviral, drug-naive, infected patients. Applying nonlinear least-squares estimation, we find that relatively small variations in parameters are capable of mimicking the highly diverse patterns found in patient viral load data. This approach yields an estimate of 2.5 days for the average lifespan of productively infected cells during primary infection, a value that is consistent with results obtained by drug perturbation experiments. We find that the data from all ten patients are consistent with a target-cell-limited model from the time of initial infection until shortly after the peak in viremia. However, the kinetics of the subsequent fall and recovery in virus concentration in some patients are not consistent with the predictions of the target-cell-limited model. We illustrate that two possible immune response mechanisms, cytotoxic T lymphocyte destruction of infected target cells and cytokine suppression of viral replication, could account for declines in viral load data not predicted by the original target-cell-limited model. We conclude that some additional process, perhaps mediated by CD8+ T cells, is important in at least some patients., (Copyright 2000 Academic Press.)

In 95% of right handed individuals the left hemisphere is dominant for speech and language function. The evidence for this is accumulated primarily from clinical populations. We investigated cortical topography of language function and lateralization in a sample of the right handed population using functional magnetic resonance imaging and two lexical-semantic paradigms. Activated cortical language networks were assessed topographically and quantitatively by using a lateralization index. As a group, we observed left hemispheric language dominance. Individually, the lateralization index varied continuously from left hemisphere dominant to bilateral representation. In males, language primarily lateralized to left, and in females, approximately half had left lateralization and the other half had bilateral representation. Our data indicate that a previous view of female bilateral hemispheric dominance for language (McGlone, 1980. Sex differences in human brain asymmetry: a critical survey. Behav Brain Sci 3:215-263; Shaywitz et al., 1995. Sex differences in the functional organization of the brain for language. Nature 373:607-609) simplifies the complexity of cortical language distribution in this population. Analysis of the distribution of the lateralization index in our study allowed us to make this difference in females apparent.

Gulf War Syndrome (GWS) is a multisystemic illness afflicting many Gulf War-era veterans. The molecular pathological basis for GWS has not been established. We sought to determine whether the presence of antibodies to squalene correlates with the presence of signs and symptoms of GWS. Participants in this blinded cohort study were individuals immunized for service in Desert Shield/Desert Storm during 1990-1991. They included 144 Gulf War-era veterans or military employees (58 in the blinded study), 48 blood donors, 40 systemic lupus erythematosus patients, 34 silicone breast implant recipients, and 30 chronic fatigue syndrome patients. Serum antibodies to squalene were measured. In our small cohort, the substantial majority (95%) of overtly ill deployed GWS patients had antibodies to squalene. All (100%) GWS patients immunized for service in Desert Shield/Desert Storm who did not deploy, but had the same signs and symptoms as those who did deploy, had antibodies to squalene. In contrast, none (0%) of the deployed Persian Gulf veterans not showing signs and symptoms of GWS have antibodies to squalene. Neither patients with idiopathic autoimmune disease nor healthy controls had detectable serum antibodies to squalene. The majority of symptomatic GWS patients had serum antibodies to squalene., (Copyright 2000 Academic Press.)

Originally, prostate-specific membrane antigen (PSMA) was described in benign and malignant prostate cells. On the basis of recent reports that this antigen also is expressed in normal renal proximal tubular cells and in the neovascular endothelium associated with renal carcinoma, we used a nested reverse transcriptase-polymerase chain reaction assay to evaluate whether PSMA-expressing cells might be present in specimens of peripheral blood obtained from renal cancer patients, benign renal tumor patients, and healthy volunteers. Our reverse transcriptase-polymerase chain reaction PSMA assay had a sensitivity of detecting 1 lymph node prostate cancer (LNCaP) per 10(7) lymphocytes. None of the 20 non-renal cancer controls were positive for PSMA mRNA, whereas 11 of 50 patients (22%) with diagnosed renal cancer were positive. Despite a comparative increase of PSMA positivity with stage, no statistical correlation was found. However, 44% of PSMA-positive patients had tumor size greater than 12 cm, versus only 9% in patients negative for PSMA (P = .03), and 67% of positive PSMA patients were found to have vascular invasion versus only 16% of patients negative for PSMA (P = .006; odds ratio, 10.8). This preliminary study suggests the possibility that PSMA expression in peripheral blood might be a useful biomarker for detecting or monitoring the progression of renal cancer in patients.

Kell blood group protein shares a consensus sequence (H.E.X.X.H) with a large family of zinc-dependent endopeptidases. Kell has closest homology with neutral endopeptidase 24.11, endothelin converting enzyme-1 (ECE-1), and the PEX gene product that, as a group, comprise the M13 subfamily of mammalian neutral endopeptidases. The proteolytic activity of the M13 members, but not of Kell, has been previously demonstrated. A secreted form of wild-type Kell protein (s-Kell), devoid of the intracellular and transmembrane domains, was expressed in sf9 cells. As a negative control, an inactive mutant Kell protein (E582G) was expressed. As determined by N-terminal amino acid sequencing and mass spectrometry of the cleaved products, wild-type s-Kell, but not the control mutant protein, specifically cleaved big endothelin-3 (ET-3) at Trp(21)-Ile(22), yielding ET-3, and, to a much lesser extent, also cleaved big ET-1 and big ET-2 at Trp(21)-Val(22), yielding ET-1 and ET-2. Enzymatic activity was partially inhibited by phosphoramidon. s-Kell has an acidic pH optimum (pH 6.0 to 6.5). Like the recombinant protein, red blood cells of common Kell phenotype also preferentially process big ET-3, in contrast to Ko (null) cells that do not. These data demonstrate that the Kell blood group protein is a proteolytic enzyme that processes big ET-3, generating ET-3, a potent bioactive peptide with multiple biological roles.

The fluorescence energy transfer (FET) between Acridine Orange and Safranine T, two intercalators of DNA, was studied in this paper. The FET efficiency between Acridine Orange and Safranine T is higher and the critical distance, R(0), is longer in the intercalated state than in the free one. A new method for the determination of calf thymus DNA (ctDNA) was presented. The linear range of the calibration curve is (0 approximately 1.1)x10(-5) mol l(-1) in bases for ctDNA, and the limit of detection is 2.6x10(-7) mol l(-1).

The integrase family of site-specific recombinases catalyzes conservative rearrangements between defined segments of DNA. A highly conserved tetrad (RHRY) of catalytic residues is essential for this process. This tetrad is dispersed in two motifs in the linear sequence, but is configured appropriately in the catalytic pocket to execute the strand cleavage and rejoining reactions. A third conserved motif has been identified in the Xer subgroup of the integrase family. Mutational analysis of 12 conserved residues in this motif in the XerD protein from Salmonella typhimurium led to the identification of an essential fifth catalytic residue (lysine 172) which is implicated in strand cleavage or exchange. This lysine residue occupies part of the turn of an antiparallel beta-hairpin which forms one side of the catalytic cleft in XerD, and is found at similar positions among evolutionarily diverse integrase family members. Related antiparallel beta-hairpins are present in eucaryotic type IB topoisomerase enzymes which also contain a critical lysine residue in the turn of the hairpin. In both the integrase family and eucaryotic type IB topoisomerases, the catalytic lysine residues are in close contact with the substrates and may play similar roles in influencing the reactivity of the phosphotyrosine intermediates formed during reactions catalyzed by both enzymes., (Copyright 1999 Academic Press.)

Bispecific monoclonal antibody (BsMab) combining two different antigen binding sites, anti-biotin and anti-HRPO paratopes, could be used as a universal immunoprobe for detecting all biotinylated macromolecules. First, a mouse hybridoma cell line secreting monospecific anti-biotin Mab was generated and characterized. Second, a quadroma cell line which could continuously secrete bsMab (anti-biotin x anti-HRPO) was developed by a nonselective microelectrofusion method. The supernatant containing bsMab was collected from tissue culture medium and purified with two affinity columns. This bsMab has comparable avidity to commercial streptavidin-HRPO when tested against biotinylated macromolecules. Compared to streptavidin, this bsMab can bind the enzyme and thus eliminate the need for chemical conjugation. This bsMab can be used as a promising immunoprobe for detecting many macromolecules bearing biotin markers, such as protein, phage, liposome and DNA in different bioassay systems.

Differential pulse polarography (DPP) was applied to measure metallothionein (MT) content in the RH-35 rat hepatoma cell. The method was sensitive in measuring MT at nanogram concentrations. The MT measurement was not interfered by small molecular weight proteins. Cellular MT content increased upon incubated in medium containing different concentrations of CdCl2 or ZnCl2. The increase was concentration-dependent. In the case of CdCl2, significant increase in cellular MT content was observed at a level as low as 0.18 mg/l. The change in MT occurred when there was no significant change in cell viability or cellular protein content. The concentration of ZnCl2 needed to induce the same amount of MT as that with 0.18 mg/l CdCl2 was 12 mg/l. At high concentrations (> 12 mg/l), ZnCl2 also caused an increase in cellular protein content. Therefore, when calculation was based on cellular protein content, ZnCl2 did not caused further increase in MT. L-ascorbic caused significant increase in cellular MT content when it was presented at levels above 100 mg/l but not at 50 mg/l. The effect of L-ascorbic acid on MT induction was not dose-dependent. At 150 mg/l, the amount of MT induced was not different from that with 100 mg/l. Thus, L-ascorbic might not directly influence cellular MT content. Cysteine at 4 and 40 mg/l did not increase cellular MT content. The results of this study showed that cellular MT content could be measured by DPP. Our finding that CdCl2 at sub-lethal concentrations can effectively induce MT suggests that MT may serve as an early warning signal prior to serve as an early warning signal prior to the occurrence of cell death.

In this paper, the DNA affinity properties of Safranine T (ST), which features a phenazinyl group, were studied. The studies indicated that ST could intercalate into the stack base pairs of DNA. Intrinsic binding constants obtained by different spectral methods were consistent within experimental errors. They were of the order of 10(4) M-1 in DNA base pairs, and the binding site size was about 7 in DNA base pairs. Studies of fluorescence quenching by anionic quenchers and melting temperature of DNA all supported the intercalative binding of ST with DNA. The experiments also showed that electrostatic binding played an important role in the interaction of ST with DNA. This research offers a new intercalation functional group to DNA-targeted drug design.

Background: Retroviruses have been implicated in the aetiology of various autoimmune diseases. We used immunoblots as a surrogate test to find out whether retroviruses play a part in the development of primary biliary cirrhosis., Methods: We did western blot tests for HIV-1 and the human intracisternal A-type particle (HIAP), on serum samples from 77 patients with primary biliary cirrhosis, 126 patients with chronic liver disease, 48 patients with systemic lupus erythematosus, and 25 healthy volunteers., Findings: HIV-1 p24 gag seroreactivity was found in 27 (35%) of 77 patients with primary biliary cirrhosis, 14 (29%) of 48 patients with systemic lupus erythematosus, 14 (50%) of 28 patients with chronic viral hepatitis, and nine (39%) of 23 patients with either primary sclerosing cholangitis or biliary atresia, compared with only one (4%) of 24 patients with alcohol-related liver disease or alpha1-antitrypsin-deficiency liver disease, and only one (4%) of 25 healthy volunteers (p=0.003). Western blot reactivity to more than two HIAP proteins was found in 37 (51%) of patients with primary biliary cirrhosis, in 28 (58%) of patients with systemic lupus erythematosus, in 15 (20%) of patients with chronic viral hepatitis, and in four (17%) of those with other biliary diseases. None of the 23 patients with either alcohol-related liver disease or alpha1-antitrypsin deficiency, and only one of the healthy controls showed the same reactivity to HIAP proteins (p<0.0001). Our results showed a strong association between HIAP seroreactivity and the detection of autoantibodies to double-stranded DNA. HIAP seroreactivity was also strongly associated with the detection of mitochondrial, nuclear, and extractable nuclear antigens., Interpretation: The HIV-1 and HIAP antibody reactivity found in patients with primary biliary cirrhosis and other biliary disorders may be attributable either to an autoimmune response to antigenically related cellular proteins or to an immune response to uncharacterised viral proteins that share antigenic determinants with these retroviruses.

Xer-mediated site-specific recombination contributes to the stability of circular chromosomes in bacteria by resolving plasmid multimers and chromosome dimers to monomers prior to cell division. Two related site-specific recombinases, XerC and XerD, each catalyse one pair of strand exchange during Xer recombination. In order to relate the recently determined structure of XerD to its function, the XerD protein was subjected to pentapeptide scanning mutagenesis, which leads to a variable five amino acid cassette being introduced randomly into the target protein. This has allowed identification of regions of XerD involved in specific DNA binding, in communicating with the partner recombinase, XerC, and in catalysis and its control. The C-terminal domain of XerD, comprising two-thirds of the protein, contains the catalytic active site and comprises ten alpha helices (alphaE to alphaN) and a beta hairpin. A flexible linker connects this domain to the N-terminal domain that comprises four alpha helices (alphaA to alphaD). Pentapeptide insertions into alphaB, alphaD, alphaG, or alphaJ interfered with DNA binding. Helices alphaG and alphaJ comprise a pseudo helix-turn-helix DNA binding motif that may provide specificity of recombinase binding. An insertion in alphaL, adjacent to an active site arginine residue, led to loss of cooperative interactions between XerC and XerD and abolished recombination activity. Other insertions close to active site residues also abolished recombination activity. Proteins with an insertion in the beta hairpin turn bound DNA, interacted cooperatively with XerC and had a phenotype that is consistent with the protein being defective in XerD catalysis. This beta hairpin appears to be highly conserved in related proteins. Insertions at a number of dispersed locations did not impair XerD catalytic activity or DNA binding, but failed to allow XerC catalysis in vivo, indicating that several sites of interaction between XerD and XerC may be important for activation of XerC catalysis by XerD., (Copyright 1997 Academic Press Limited.)

Fib420 is a recently identified subclass of normal human fibrinogen in which two extended alpha chain isoforms (alphaE) replace the common alpha chains, yielding a molecule (ca. 420 kD) which is larger than the more abundant 340-kD form. Evidence for preservation of this subclass throughout vertebrate evolution suggests it performs some as yet unidentified vital function. A survey was undertaken to establish the range of plasma Fib420 levels in normal, healthy adults and in placental cord (fetal) blood. For measuring Fib420, a quantitative Western blot assay was developed using monoclonal antibody against the exon-VI encoded C-terminus of the molecule's unique alphaE chain. This alphaE chain signal was normalized to that of the beta chain, common to both fibrinogen forms. Analysis of plasma samples from the adult and newborn cohorts (n = 25 each; total fibrinogen ca. 2.6 mg/mL in both) revealed a statistically significant difference, with a mean level of 100 +/- 28 microg/mL in the neonate compared to 34 +/- 7 microg/mL in the adult. On average, 1 out of every 100 fibrinogen molecules in adult plasma belongs to the Fib420 subclass. Unlike in the newborn, adult Fib420 levels remained the same over a wide range of total plasma fibrinogen. The striking difference observed between these two cohorts suggests a changing developmental expression of the Fib420 subclass and a homeostatic control operating in later stages of life.