Hypothyroidism Modeling in Wistar Rats: Comparison of Two Surgical Methods.

In this study we present 2 surgical models of hypothyroidism in male Wistar rats (n=80) based on total thyroidectomy. Animals weighing 180-200 g were randomly divided into sham-operated group and 2 experimental groups. Thyroidectomy was performed by 2 different methods: primary ligation of either thyroid artery (TE-I) or vein (TE-II). The success of the model was verified through general postoperative conditions, serum hormone levels, histological study, and neck ultrasound. Hypothyroidism was successfully reproduced in both TE-I and TE-II models. TE-I was characterized by lower intra- and post-operative mortality, while TE-II provided better surgical exposure to the key anatomical sites.

Pleiotropic loci underlying bone mineral density and bone size identified by a bivariate genome-wide association analysis.

Aiming to identify pleiotropic genomic loci for bone mineral density and bone size, we performed a bivariate GWAS in five discovery samples and replicated in two large-scale samples. We identified 2 novel loci at 2q37.1 and 6q26. Our findings provide insight into common genetic architecture underlying both traits. INTRODUCTION: Bone mineral density (BMD) and bone size (BS) are two important factors that contribute to the development of osteoporosis and osteoporotic fracture. Both BMD and BS are highly heritable and they are genetically correlated. In this study, we aim to identify pleiotropic loci associated with BMD and BS. METHODS: We conducted a bivariate genome-wide association (GWA) analysis of hip BMD and hip BS in 6180 participants from 5 samples, followed by in silico replication in the UK Biobank study of BMD (N = 426,824) and the deCODE study of BS (N = 28,954), respectively. RESULTS: SNPs from 2 genomic loci were significant at the genome-wide significance (GWS) level (p lt; 5 × 10-8) in the discovery samples and were successfully replicated in the replication samples (2q37.1, lead SNP rs7575512, discovery p = 1.49 × 10-10, replication p = 0.05; 6q26, lead SNP rs1040724, discovery p = 1.95 × 10-8, replication p = 0.03). Functional annotations suggested functional relevance of the identified variants to bone development. CONCLUSION: Our findings provide insight into the common genetic architecture underlying BMD and BS, and enhance our understanding of the potential mechanism of osteoporosis fracture.

A novel automatic molecular test for detection of multidrug resistance tuberculosis in sputum specimen: A case control study.

MiniLab tuberculosis (ML TB) assay is a new automatic diagnostic tool for diagnosis of multidrug resistance tuberculosis (MDR-TB). This study was conducted with aims to know the performance of this assay. Sputum sample from 224 TB suspects was collected from tuberculosis suspects seeking medical care at Beijing Chest hospital. The sputum samples were directly used for smear and ML TB test. The left sputum sample was used to conduct Xpert MTB/RIF, Bactec MGIT culture and drug susceptibility test (DST). All discrepancies between the results from DST, molecular and phenotypic methods were confirmed by DNA Sequencing. The sensitivity and specificity of ML TB test for detecting MTBC from TB suspects were 95.1% and 88.9%, respectively. The sensitivity for smear negative TB suspects was 64.3%. For detection of RIF resistance, the sensitivity and specificity of ML TB test were 89.2% and 95.7%, respectively. For detection of INH resistance, the sensitivity and specificity of ML TB test were 78.3% and 98.1%, respectively. ML TB test showed similar performance to Xpert MTB/RIF for detection of MTBC and RIF resistance. In addition, ML TB also had good performance for INH resistance detection.

Association of 3q13.32 variants with hip trochanter and intertrochanter bone mineral density identified by a genome-wide association study.

We performed a GWAS of trochanter and intertrochanter bone mineral density (BMD) in the Framingham Heart Study and replicated in three independent studies. Our results identified one novel locus around the associated variations at chromosomal region 3q13.32 and replicated two loci at chromosomal regions 3p21 and 8q24. Our findings provide useful insights that enhance our understanding of bone development, osteoporosis, and fracture pathogenesis. INTRODUCTION: Hip trochanter (TRO) and intertrochanter (INT) subregions have important clinical relevance to subtrochanteric and intertrochanteric fractures but have rarely been studied by genome-wide association studies (GWASs). METHODS: Aiming to identify genomic loci associated with BMD variation at TRO and INT regions, we performed a GWAS utilizing the Framingham Heart Study (FHS, N = 6,912) as discovery sample and utilized the Women's Health Initiative (WHI) African-American subsample (N = 845), WHI Hispanic subsample (N = 446), and Omaha osteoporosis study (N = 971), for replication. RESULTS: Combining the evidence from both the discovery and the replication samples, we identified one novel locus around the associated variations at chromosomal region 3q13.32 (rs1949542, discovery p = 6.16 × 10-8, replication p = 2.86 × 10-4 for INT-BMD; discovery p = 1.35 × 10-7, replication p = 4.16 × 10-4 for TRO-BMD, closest gene RP11-384F7.1). We also replicated two loci at chromosomal regions 3p21 (rs148725943, discovery p = 6.61 × 10-7, replication p = 5.22 × 10-4 for TRO-BMD, closest gene CTNNB1) and 8q24 (rs7839059, discovery p = 2.28 × 10-7, replication p = 1.55 × 10-3 for TRO-BMD, closest gene TNFRSF11B) that were reported previously. We demonstrated that the effects at both 3q13.32 and 3p21 were specific to the TRO, but not to the femoral neck and spine. In contrast, the effect at 8q24 was common to all the sites. CONCLUSION: Our findings provide useful insights that enhance our understanding of bone development, osteoporosis, and fracture pathogenesis.

Hemagglutinin stalk-based universal vaccine constructs protect against group 2 influenza A viruses.

Current influenza virus vaccines contain H1N1 (phylogenetic group 1 hemagglutinin), H3N2 (phylogenetic group 2 hemagglutinin), and influenza B virus components. These vaccines induce good protection against closely matched strains by predominantly eliciting antibodies against the membrane distal globular head domain of their respective viral hemagglutinins. This domain, however, undergoes rapid antigenic drift, allowing the virus to escape neutralizing antibody responses. The membrane proximal stalk domain of the hemagglutinin is much more conserved compared to the head domain. In recent years, a growing collection of antibodies that neutralize a broad range of influenza virus strains and subtypes by binding to this domain has been isolated. Here, we demonstrate that a vaccination strategy based on the stalk domain of the H3 hemagglutinin (group 2) induces in mice broadly neutralizing anti-stalk antibodies that are highly cross-reactive to heterologous H3, H10, H14, H15, and H7 (derived from the novel Chinese H7N9 virus) hemagglutinins. Furthermore, we demonstrate that these antibodies confer broad protection against influenza viruses expressing various group 2 hemagglutinins, including an H7 subtype. Through passive transfer experiments, we show that the protection is mediated mainly by neutralizing antibodies against the stalk domain. Our data suggest that, in mice, a vaccine strategy based on the hemagglutinin stalk domain can protect against viruses expressing divergent group 2 hemagglutinins.

A Sendai virus-derived RNA agonist of RIG-I as a virus vaccine adjuvant.

The innate immune system is responsible for recognizing invading pathogens and initiating a protective response. In particular, the retinoic acid-inducible gene 1 protein (RIG-I) participates in the recognition of single- and double-stranded RNA viruses. RIG-I activation leads to the production of an appropriate cytokine and chemokine cocktail that stimulates an antiviral state and drives the adaptive immune system toward an efficient and specific response against the ongoing infection. One of the best-characterized natural RIG-I agonists is the defective interfering (DI) RNA produced by Sendai virus strain Cantell. This 546-nucleotide RNA is a well-known activator of the innate immune system and an extremely potent inducer of type I interferon. We designed an in vitro-transcribed RNA that retains the type I interferon stimulatory properties, and the RIG-I affinity of the Sendai virus produced DI RNA both in vitro and in vivo. This in vitro-synthesized RNA is capable of enhancing the production of anti-influenza virus hemagglutinin (HA)-specific IgG after intramuscular or intranasal coadministration with inactivated H1N1 2009 pandemic vaccine. Furthermore, our adjuvant is equally effective at increasing the efficiency of an influenza A/Puerto Rico/8/34 virus inactivated vaccine as a poly(I·C)- or a squalene-based adjuvant. Our in vitro-transcribed DI RNA represents an excellent tool for the study of RIG-I agonists as vaccine adjuvants and a starting point in the development of such a vaccine.

Assessing validity of the adapted Arabic Paediatric Asthma Quality of Life Questionnaire among Egyptian children with asthma.

The recent focus in asthma management is rendering children a better quality of life (QOL). Validity and reliability of an adapted Arabic translation of the Paediatric Asthma Quality of Life Questionnaire (PAQLQ-A) among Egyptians was assessed in a cohort of 103 asthmatic children aged 8-16 years. Discriminative validity of mean scores was significantly higher among mild asthmatics than those with moderate/severe asthma. Construct validity of domains was significantly negatively correlated with clinical severity score. Reliability and internal consistency were assessed using Cronbach alpha coefficient (alpha = 0.84). Reproducibility and responsiveness were high among both stable and unstable asthma patients. PAQLQ-A is valid and reliable for assessing QOL among Egyptian asthmatic children.

Female youth health promotion model in primary health care: a community-based study in rural upper Egypt.

The health status of underprivileged young females is a global concern. This intervention study in rural Upper Egypt used an integrated approach to develop a model for primary care health promotion services to female adolescents. An initial household survey and focus group discussions identified the health problems of a sample of 671 adolescent women aged 12-20 years recruited from one village. Interventions included training courses for health care providers on relevant health topics and on client-provider interaction skills; community and local authority mobilization; and health education sessions and a special record system for the women. An increase was seen in the utilization of primary care services.

HLA-DPB1, -DRB1, and -DQB1 polymorphism defined in Ewenki ethnic minority of China Inner Mongolia Autonomous Region.

In the present study, DNA typing for human leucocyte antigen (HLA)-DPB1, -DRB1, and -DQB1 was performed using polymerase chain reaction-sequence-based-typing (PCR-SBT) method on 94 randomly selected, healthy, unrelated individuals from the Ewenki ethnic population in Inner Mongolia Autonomous Region of China. A total of 64 alleles: 25 in DRB1, 19 in DQB1 and 20 in DPB1, were found. Among the 25 detected DRB1 alleles, DRB1*090102, DRB1*030101, DRB1*040101, DRB1*070101, and DRB1*120101/1206 were commonly observed, with frequencies of 16.0%, 13.3%, 10.1%, 7.4%, and 7.4%, respectively. The most predominant DQB1 allele was DQB1*030101/0309 with the frequency of 27.7%, followed by DQB1*0201/0202 (19.7%), DQB1*030302 (12.8%), DQB1*060101/060103 (6.4%), and DQB1*050201 (5.9%). Of the 20 detected DPB1 alleles, DPB1*020102 was the most frequent allele with the frequency of 25.5%. DPB1*0402 (21.3%), DPB1*0401 (20.2%), DPB1*0501 (10.6%) and DPB1*4101 (3.7%) were also very frequent alleles. The most frequent two-locus haplotypes observed in the Ewenki were: DRB1*030101-DQB1*0201/0202(10.7%), DRB1*090102-DQB1*03032(9.8%), DRB1*070101-DQB1*0201/0202 (5.5%), DRB1*070101-DQB1*030302 (5.2%) and DRB1*120101/1206-DQB1*030101/0309 (4.6%). The distribution of the HLA class II alleles and haplotypes frequencies as well as the dendrogram showed that the Ewenki population belongs to the northern group of Chinese.

Beyond information retrieval--medical question answering.

Physicians have many questions when caring for patients, and frequently need to seek answers for their questions. Information retrieval systems (e.g., PubMed) typically return a list of documents in response to a user's query. Frequently the number of returned documents is large and makes physicians' information seeking "practical only 'after hours' and not in the clinical settings". Question answering techniques are based on automatically analyzing thousands of electronic documents to generate short-text answers in response to clinical questions that are posed by physicians. The authors address physicians' information needs and described the design, implementation, and evaluation of the medical question answering system (MedQA). Although our long term goal is to enable MedQA to answer all types of medical questions, currently, we implemented MedQA to integrate information retrieval, extraction, and summarization techniques to automatically generate paragraph-level text for definitional questions (i.e., "What is X?"). MedQA can be accessed at http://www.dbmi.columbia.edu/~yuh9001/research/MedQA.html.

[Study on the application of random amplified polymorphic DNA in Yersinia pestis genotyping].

OBJECTIVE: To study the type of 103 Y. pestis strains that isolated from different plague foci and different ecological types. METHOD: Random amplified polymorphic DNA (RAPD) was used. RESULTS: Y. pestis were divided into two RAPD types: most strains that isolated from elsewhere in the country belonged to RAPD-1 type, while most strains that isolated from Qinghai province were RAPD-2 type. CONCLUSION: The genome structures of different ecotype Y. pestis were different providing the foundation of further research and prevention of plague.

[The sequence analysis of the 3' end of intron 15 from human LDL receptor gene and its application in the study of gene diagnosis for familial hypercholesterolaemia].

To understand intron 15 of human LDL receptor gene, the DNA fragments from exon 15 to exon 16 and the 3' end of intron 15 were amplified with long chain PCR and anchored PCR. The 3' end of intron 15 was sequenced with Dynalbeads-Streptavidin Solid Phase technique. The sequence analysis showed that the 3' end of intron 15 contained the 3' splicing site and the branch site at 31 nucleotides upstream of the 3' end. Besides the authentic branch site, it is possible that the 3' end of intron 15 contains a cryptic site (GCCTCAC) at 20 nucleotides upstream of the 3' end. The sequences suggest that the PvuII polymorphism at Intron 15 is caused by the T-C substitution. According to the sequences of the 3' end, the new PCR-RFLP protocol for detection of PvuII polymorphism at intron 15 was developed. Using this protocol a representative familial hypercholesterolaemia family was identified with linkage analysis of PvuII polymorphism at intron 15.