Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol for bioluminescent optical imaging of HO-1-luciferase transgenic mice reported in this manuscript provides a rapid in vivo assay of the upregulation of the heme oxygenase-1 (HO-1) gene expression in response to the Gua Sha procedure. HO-1 has long been known to provide cytoprotection against oxidative stress. The upregulation of HO-1, assessed by the bioluminescence output, is thought to represent an antioxidative response to circulating hemoglobin products released by Gua Sha. Gua Sha was administered by repeated strokes of a smooth spoon edge over lubricated skin on the back or other targeted body part of the transgenic mouse until petechiae (splinter hemorrhages) or ecchymosis (bruises) indicative of extravasation of blood from subcutaneous capillaries was observed. After Gua Sha, bioluminescence imaging sessions were carried out daily for several days to follow the dynamics of HO-1 expression in multiple internal organs

Disease Burden of Clostridium difficile Infections in Adults, Hong Kong, China, 2006-2014

Cross-sectional studies suggest an increasing trend in incidence and relatively low recurrence rates of Clostridium difficile infections in Asia than in Europe and North America. The temporal trend of C. difficile infection in Asia is not completely understood. We conducted a territory-wide population-based observational study to investigate the burden and clinical outcomes in Hong Kong, China, over a 9-year period. A total of 15,753 cases were identified, including 14,402 (91.4%) healthcare-associated cases and 817 (5.1%) community-associated cases. After adjustment for diagnostic test, we found that incidence increased from 15.41 cases/100,000 persons in 2006 to 36.31 cases/100,000 persons in 2014, an annual increase of 26%. This increase was associated with elderly patients, for whom incidence increased 3-fold over the period. Recurrence at 60 days increased from 5.7% in 2006 to 9.1% in 2014 (p<0.001). Our data suggest the need for further surveillance, especially in Asia, which contains ≈60% of the world’s population

Wild Type and Mutant 2009 Pandemic Influenza A (H1N1) Viruses Cause More Severe Disease and Higher Mortality in Pregnant BALB/c Mice

BACKGROUND: Pregnant women infected by the pandemic influenza A (H1N1) 2009 virus had more severe disease and higher mortality but its pathogenesis is still unclear. PRINCIPAL FINDINGS: We showed that higher mortality, more severe pneumonitis, higher pulmonary viral load, lower peripheral blood T lymphocytes and antibody responses, higher levels of proinflammatory cytokines and chemokines, and worse fetal development occurred in pregnant mice than non-pregnant controls infected by either wild type (clinical isolate) or mouse-adapted mutant virus with D222G substitution in hemagglutinin. These disease-associated changes and the lower respiratory tract involvement were worse in pregnant mice challenged by mutant virus. Though human placental origin JEG-3 cell line could be infected and proinflammatory cytokines or chemokines were elevated in amniotic fluid of some mice, no placental or fetal involvement by virus were detected by culture, real-time reverse transcription polymerase chain reaction or histopathological changes. Dual immunofluorescent staining of viral nucleoprotein and type II alveolar cell marker SP-C protein suggested that the majority of infected alveolar epithelial cells were type II pneumocytes. CONCLUSION: The adverse effect of this pandemic virus on maternal and fetal outcome is largely related to the severe pulmonary disease and the indirect effect of inflammatory cytokine spillover into the systemic circulation

Multiple Gene Polymorphisms in the Complement Factor H Gene Are Associated with Exudative Age-Related Macular Degeneration in Chinese

PURPOSE. Variants in the complement factor H (CFH) gene have been shown to be strongly associated with age-related macular degeneration (AMD). In this study, sequence alterations in CFH were investigated in 163 Chinese patients with exudative AMD and 155 unrelated Chinese control subjects. METHODS. All the 22 CFH exons, intron-exon boundaries, and promoter sequences were screened by polymerase chain reaction and DNA sequencing. RESULTS. Fifty-eight sequence changes, 42 of them novel, were identified. Six SNPs with an allele frequency Ͼ30% were significantly associated with exudative AMD. SNP rs3753396 was novel; the rest had been reported: rs3753394, rs551397, rs800292, rs2274700, and rs1329428. Two haplotype blocks were constructed. The TG haplotype for rs551397 and rs800292 was the major haplotype that conferred a significantly increased susceptibility to exudative AMD (P corr ϭ 0.0001, OR ϭ 1.91, 95% CI ϭ 1.36 -2.68). CONCLUSIONS. The findings support prior evidence that the CFH gene is one of the AMD-associated genes. There is a different distribution pattern of CFH variants in the Chinese compared with other populations. Individual SNP and haplotype analyses revealed that the ancient alleles at the 5Ј end of CFH contribute to an increased susceptibility to exudative AMD. (Invest Ophthalmol Vis Sci. 2008;49:3312-3317) DOI:10.1167/iovs.07-1517 A ge-related macular degeneration (AMD; MIM 603075; Mendelian Inheritance in Man) is a major cause of irreversible visual impairment and blindness in people older than 65 years in developed countries. 1,2 The occurrence of AMD is pan ethnic, and a high prevalence AMD has been reported in the elderly Chinese population. 5 Therefore, a greater understanding of the primary pathophysiology is needed to advance treatment and preventive measures. The etiology of AMD is complex and multifactorial, probably resulting from interactions between environmental and multigenetic factors. 6 Genetic association studies have revealed that single nucleotide polymorphisms (SNPs) in the complement factor H gene (CFH; MIM 134370; e.g., Tyr402His) are significantly associated with susceptibility to AMD. 25 A fine-scale linkage disequilibrium mapping of AMD in the CFH region detected a point location of a causal variant between exons 1 and 2 of CFH other than exon 9 for Tyr402His. MATERIALS AND METHODS Patients and Control Subjects 21 Also recruited and given complete ophthalmic examinations were 155 unrelated control subjects, 72 men and 83 women ranging in age at recruitment from 60 to 99 years (mean Ϯ SD, 73.1 Ϯ 6.5 years). They matched the patients by age and gender and had no sign of AMD or other eye diseases, except mild myopia or senile cataract. The study protocol was approved by the Ethics Committee on Human Research, the Chinese University of Hong Kong. All the procedures used conformed to the tenets of the Declaration of Helsinki. Informed consent was obtained from all study subjects after explanation of the nature of the study. Sample Collection, PCR Amplification, DNA Sequencing, and SNP Genotyping Venous blood was obtained from each study subject, and genomic DNA was extracted with a DNA blood kit (QIAamp; Qiagen, Hilden, Germany). The promoter sequence up to Ϫ867 upstream and all coding sequences of the CFH gene, including intron-exon boundaries, were screened for sequence alterations. Primers were generated based on the GenBank sequence of CFH (NM_000186.2; http://www.ncbi. nlm.nih.gov/Genbank; provided in the public domain by the National Center for Biotechnology Information, Bethesda, MD). PCR was performed on a thermal cycler (model 9700; Applied Biosystems, Inc. [ABI], Foster City, CA) with optimized protocols 27 Statistical Analysis Hardy-Weinberg equilibrium (HWE) for each polymorphism was tested by 2 test. Allele or genotype frequencies between cases and control subjects were compared by 2 analysis or the Fisher exact test. The odds ratios (ORs) of the alleles and haplotypes were estimated by 2 test (SPSS ver.15.0; SPSS Inc., Chicago, IL). Population attributable risk (PAR) of the risk genotype was calculated with the formula f(R Ϫ 1)/R, where f is the faction of cases with the risk genotype and R is the measure of OR 8 . A pair-wise linkage disequilibrium (LD, DЈ) estimation between polymorphisms with a minor allele frequency (MAF) Ͼ 1%, and EM-based haplotype association analysis were performed with Haploview (ver. 3.32, from http://www.broad.mit.edu/mpg/ haploview/ provided in the public domain by the Broad Institute, Massachusetts Institute of Technology, Cambridge, MA). For multiple comparison, probabilities were corrected by permutation test (iterations, 10,000). Statistical significance was defined as a corrected P (P corr ) Ͻ 0.05. RESULTS CFH Variants in the Study Subjects A total of 58 sequence variations were identified, all of which followed Hardy Weinberg Equilibrium Six of the seven common variants Six SNPs were identified in the promoter, all supported decreased susceptibility to AMD Haplotype Association Analysis LD analysis revealed extension of LD throughout the CFH gene. We included SNPs with MAF Ͼ 5% and two missense changes, rs1061170 (Tyr402His) and Val837Ile, in our haplotype association analysis. Two distinct haplotype blocks were detected The haplotypes H3 and H4, which were defined by all six AMD-associated SNPs, conferred significantly reduced or increased AMD susceptibility (H3: OR ϭ 0.56, 95% CI ϭ 0.39 -0.80; H4: OR ϭ 1.63, 95% CI ϭ 1.19 -2.23). When a G allele of rs1065489 (Asp936Glu) was included in these two haplotypes, the H5, which contained all the alleles in H3, remain significantly associated with the disease (P corr ϭ 0.0012). However, when a G allele or a T allele was added to the H4, the newly constructed H6 and H7 were no longer AMD associated (P corr ϭ 0.052 and 0.177, respectively). We constructed two-allele haplotypes by using rs800292 (Val62Ile) with the uncommon SNPs rs1061170 (Tyr402His) and Val837Ile, to investigate the effects of the minor variants. H10 and H11, containing a T allele of rs1061170 (Tyr402His), remained significantly associated with AMD. However, the haplotypes containing a C allele of rs1061170 DISCUSSION Although the pathogenesis of exudative AMD has not been definitively elucidated, studies in the past few years have revealed important information on its genetic basis. Polymorphisms in the CFH gene have been shown to be AMD associated in different ethnic groups, although there are obvious differences in the occurrence of disease-susceptible SNPs between Caucasian and Oriental populations. 26 mapped a point location for a causal variant between exons 1 and 2, which approximates block 1 in our present study, suggesting that the 5Ј region of the CFH (N-terminal of factor H) is commonly associated with AMD in both Chinese and Caucasians. We found haplotype block 2 spanning a region from exon 10 to intron 15 and containing SNP rs2274700 (Ala473Ala, exon 10), which have recently been shown to have a strong association with AMD in Caucasians and Japanese. Besides the haplotypes in the two haplotype blocks, the haplotypes defined by the six common SNPs (H3, H4) were also significantly associated with exudative AMD. However, when Asp936Glu (in exon 18) was included in the at-risk haplotype H4 for association analysis, the haplotypes H6 and H7, including a G or a T allele respectively, were no longer significantly associated with the disease (P corr Ͼ 0.05). Thus, Asp936Glu is less likely to be a risk factor for exudative AMD in Chinese individuals, indicating the C-terminal of the factor H contributes less than other parts of the polypeptide to the development of exudative AMD. This observation is consistent with the findings of Hageman et al

Impact of genetic loci identified in genome-wide association studies on diabetic retinopathy in Chinese patients with type 2 diabetes

© 2016, Association for Research in Vision and Ophthalmology Inc. All rights reserved.PURPOSE. Diabetic retinopathy (DR) is a common microvascular complication of type 2 diabetes (T2DM). Genome-wide association studies (GWAS) had identified novel DRsusceptibility genetic variants in various populations. We examined the associations of these DR-associated single nucleotide polymorphisms (SNPs) with severe DR in a Chinese T2DM cohort. METHODS. Cross-sectional case-control studies on sight-threatening DR (STDR) and proliferative DR (PDR) were performed. We genotyped 38 SNPs showing top association signals with DR in previous GWAS in 567 STDR cases, including 309 with PDR and 1490 non-DR controls. Multiple logistic regression models with adjustment for conventional risk factors, including age, sex, duration of diabetes, and presence of hypertension, were employed. RESULTS. The strongest association was found at INSR rs2115386, an intronic SNP of INSR: Padjusted = 9.13 × 10-4 (odds ratio [OR],1.28; 95% confidence interval [95%CI], 1.11-1.48) for STDR, and Padjusted = 1.12 × 10-4 (OR [95%CI],1.44 [1.20-1.74]) for PDR. rs599019 located downstream of COLEC12 (Padjusted = 0.019; OR [95%CI],1.19 [1.03-1.38]) and rs4462262 located at an intergenic region between ZWINT and MRPS35P3 (Padjusted = 0.041; OR [95%CI],1.38[1.01-1.89]) also were significantly associated with STDR, but not with PDR alone. On the other hand, MYT1L-LOC729897 rs10199521 (Padjusted = 0.022; OR [95%CI],1.25 [1.03-1.51]) and API5 rs899036 (Padjusted = 0.049; OR [95%CI],1.36 [1.00-1.85]) showed significant independent associations only with PDR. Similar results were obtained when hemoglobin A1c also was included in the adjustment models. CONCLUSIONS. We demonstrated the significant and independent associations of several GWAS-identified SNPs with DR in Chinese T2DM patients with severe DR. The findings on INSR rs2115386 are supportive of the role of insulin resistance, or the compensatory hyperinsulinemia, in the pathogenesis of DR.Link_to_subscribed_fulltex

Identification of a Functional Genetic Variant at 16q12.1 for Breast Cancer Risk: Results from the Asia Breast Cancer Consortium

Genetic factors play an important role in the etiology of breast cancer. We carried out a multi-stage genome-wide association (GWA) study in over 28,000 cases and controls recruited from 12 studies conducted in Asian and European American women to identify genetic susceptibility loci for breast cancer. After analyzing 684,457 SNPs in 2,073 cases and 2,084 controls in Chinese women, we evaluated 53 SNPs for fast-track replication in an independent set of 4,425 cases and 1,915 controls of Chinese origin. Four replicated SNPs were further investigated in an independent set of 6,173 cases and 6,340 controls from seven other studies conducted in Asian women. SNP rs4784227 was consistently associated with breast cancer risk across all studies with adjusted odds ratios (95% confidence intervals) of 1.25 (1.20−1.31) per allele (P = 3.2×10−25) in the pooled analysis of samples from all Asian samples. This SNP was also associated with breast cancer risk among European Americans (per allele OR  = 1.19, 95% CI  = 1.09−1.31, P = 1.3×10−4, 2,797 cases and 2,662 controls). SNP rs4784227 is located at 16q12.1, a region identified previously for breast cancer risk among Europeans. The association of this SNP with breast cancer risk remained highly statistically significant in Asians after adjusting for previously-reported SNPs in this region. In vitro experiments using both luciferase reporter and electrophoretic mobility shift assays demonstrated functional significance of this SNP. These results provide strong evidence implicating rs4784227 as a functional causal variant for breast cancer in the locus 16q12.1 and demonstrate the utility of conducting genetic association studies in populations with different genetic architectures

RNA-Seq Analyses Generate Comprehensive Transcriptomic Landscape and Reveal Complex Transcript Patterns in Hepatocellular Carcinoma

RNA-seq is a powerful tool for comprehensive characterization of whole transcriptome at both gene and exon levels and with a unique ability of identifying novel splicing variants. To date, RNA-seq analysis of HBV-related hepatocellular carcinoma (HCC) has not been reported. In this study, we performed transcriptome analyses for 10 matched pairs of cancer and non-cancerous tissues from HCC patients on Solexa/Illumina GAII platform. On average, about 21.6 million sequencing reads and 10.6 million aligned reads were obtained for samples sequenced on each lane, which was able to identify >50% of all the annotated genes for each sample. Furthermore, we identified 1,378 significantly differently expressed genes (DEGs) and 24, 338 differentially expressed exons (DEEs). Comprehensive function analyses indicated that cell growth-related, metabolism-related and immune-related pathways were most significantly enriched by DEGs, pointing to a complex mechanism for HCC carcinogenesis. Positional gene enrichment analysis showed that DEGs were most significantly enriched at chromosome 8q21.3–24.3. The most interesting findings were from the analysis at exon levels where we characterized three major patterns of expression changes between gene and exon levels, implying a much complex landscape of transcript-specific differential expressions in HCC. Finally, we identified a novel highly up-regulated exon-exon junction in ATAD2 gene in HCC tissues. Overall, to our best knowledge, our study represents the most comprehensive characterization of HBV-related HCC transcriptome including exon level expression changes and novel splicing variants, which illustrated the power of RNA-seq and provided important clues for understanding the molecular mechanisms of HCC pathogenesis at system-wide levels

Phase-specific and lifetime costs of cancer care in Ontario, Canada

BACKGROUND: Cancer is a major public health issue and represents a significant economic burden to health care systems worldwide. The objective of this analysis was to estimate phase-specific, 5-year and lifetime net costs for the 21 most prevalent cancer sites, and remaining tumour sites combined, in Ontario, Canada. METHODS: We selected all adult patients diagnosed with a primary cancer between 1997 and 2007, with valid ICD-O site and histology codes, and who survived 30 days or more after diagnosis, from the Ontario Cancer Registry (N = 394,092). Patients were linked to treatment data from Cancer Care Ontario and administrative health care databases at the Institute for Clinical and Evaluative Sciences. Net costs (i.e., cost difference between patients and matched non-cancer control subjects) were estimated by phase of care and sex, and used to estimate 5-year and lifetime costs. RESULTS: Mean net costs of care (2009 CAD) were highest in the initial (6 months post-diagnosis) and terminal (12 months pre-death) phases, and lowest in the (3 months) pre-diagnosis and continuing phases of care. Phase-specific net costs were generally lowest for melanoma and highest for brain cancer. Mean 5-year net costs varied from less than $25,000 for melanoma, thyroid and testicular cancers to more than$60,000 for multiple myeloma and leukemia. Lifetime costs ranged from less than $55,000 for lung and liver cancers to over$110,000 for leukemia, multiple myeloma, lymphoma and breast cancer. CONCLUSIONS: Costs of cancer care are substantial and vary by cancer site, phase of care and time horizon analyzed. These cost estimates are valuable to decision makers to understand the economic burden of cancer care and may be useful inputs to researchers undertaking cancer-related economic evaluations