Plasma Metabonomics of Human Adenovirus-infected Patients with Pneumonia and Upper Respiratory Tract Infection

Objective Human adenovirus(HAdV)infection is common and can develop to serious conditions with high mortality,yet the mechanism of HAdV infection remains unclear.In the present study,the serum metabolite profiles of HAdV-7-infected patients with pneumonia or upper respiratory tract infection(URTI)were explored.Methods In total,35 patients were enrolled in the study following an outbreak of HAdV-7 in the army,of whom 14 had pneumonia and 21 had URTI.Blood samples were collected at the acute stage and at the recovery stage and were analyzed by untargeted metabolomics.Results Over 90% of the differential metabolites identified between the pneumonia patients and URTI patients were lipids and lipid-like molecules,including glycerophospholipids,fatty acyls,and sphingolipids.The metabolic pathways that were significantly enriched were primarily the lipid metabolism pathways,including sphingolipid metabolism,glycerophospholipid metabolism,and linoleic acid metabolism.The sphingolipid metabolism was identified as a significantly differential pathway between the pneumonia patients and URTI patients and between the acute and recovery stages for the pneumonia patients,but not between the acute and recovery stages for the URTI patients.Ceramide and lactosylceramide,involved in sphingolipid metabolism,were significantly higher in the pneumonia patients than in the URTI patients with good discrimination abilities[area under curve(AUC)0.742 and 0.716,respectively;combination AUC 0.801].Conclusion Our results suggested that HAdV modulated lipid metabolism for both the patients with URTI and pneumonia,especially the sphingolipid metabolism involving ceramide and lactosylceramide,which might thus be a potential intervention target in the treatment of HAdV infection.

Impact of host genetics on gut microbiome: Take-home lessons from human and mouse studies

The intestinal microbiome has emerged as an important component involved in various diseases.Therefore,the interest in understanding the factors shaping its composition is growing.The gut microbiome,often defined as a complex trait,contains diverse components and its properties are determined by a combination of external and internal effects.Although much effort has been invested so far,it is still difficult to evaluate the extent to which human genetics shape the composition of the gut microbiota.However,in mouse studies,where the environmental factors are better controlled,the effect of the genetic background was significant.The purpose of this paper is to provide a current assessment of the role of human host genetics in shaping the gut microbiome composition.Despite the inconsistency of the reported results,it can be estimated that the genetic factor affects a portion of the microbiome.However,this effect is currently lower than the initial estimates,and it is difficult to separate the genetic influence from the environmental effect.Additionally,despite the differences between the microbial composition of humans and mice,results from mouse models can strengthen our knowledge of host genetics underlying the human gut microbial variation.

Phylogenetic analysis of the genomes of two strains of human adenovirus type 3

Human adenovirus type 3 （HAdV-3） is widely prevalent all over the world, especially in Asia. The objective of this study is to carry out complete genomic DNA sequencing and the phylogenetic analysis for two strains （Guangzhou01 and Guangzhou02） of HAdV-3 wild virus isolated from South China. Nasopharyngeal secretion aspirate specimens of sick children were inoculated into HEp-2 and HeLa culture tubes, and the cultures were identified by neutralization assay with type-specific reference rabbit antiserum. Type-specific primers were also utilized to confirm the serotype. The restriction fragments of HAdV genome DNA were cloned into pBlueScript SK （ ＋ ） vectors and sequenced, and the 5＇ and 3＇ ends of the linear HAdV-3 genome were directly sequenced with double purified genomic DNA as templates. General features of the HAdV-3 genome sequences were explored by using several bio-software. Phylogenetic analysis was done with MEGA 3.0 software. The genomic sequences of Guangzhou01 and Guangzhou02 possess the same 4 early regions and 5 late regions and have 39 coding sequences and two RNA coding sequences. Other non-coding regions are conservative. Inverted repeats and palindromes were identified in the genome sequences. The genomes of group B human adenovirus as well as HAdV-3 have close phylogenetic relationship with that of chimpanzee adenovirus type 21. The genomic lengths of these two isolated strains are 35 273 bp and 35 269 bp, respectively. The phylogenetic analysis showed that HAdV-B species has some relationship with certain types of chimpanzee adenovirus.

Molecular Characterization of Human Respiratory Adenovirus Infection in Children from November 2016 to October 2017 in Xining City, China

Human adenoviruses (HAdVs) belong to the genus Mastadenovirus, family Adenoviridae, and can cause respiratory tract and gastrointestinal tract infections, as well as ocular infections in humans[1]. Till date, a total of 84 unique genotypes of AdVs have been identified and classified as human Mastadenovirus species A to G, and specific types are often associated with certain clinical symptoms, epidemiological settings, and demographic risk groups[2]. Among these species, members of the species HAdV-B (HAdV-3, HAdV-7, HAdV-11, HAdV-16, HAdV-21, HAdV-34, HAdV-35, HAdV-50, etc), HAdV-C (HAdV-1, HAdV-2, HAdV-5, and HAdV-6), and HAdV-E (HAdV-4) have been generally associated with respiratory infections[3]. HAdV infections are mild and self-limited in healthy individuals, whereas they can result in high mortality rates in immunocompetent and immunocompromized patients[4].

Genetic Correction and Hepatic Differentiation of Hemophilia B-specific Human Induced Pluripotent Stem Cells

Objective To genetically correct a disease-causing point mutation in human induced pluripotent stem cells （iPSCs） derived from a hemophilia B patient. Methods First, the disease-causing mutation was detected by sequencing the encoding area of human coagulation factor IX （F IX） gene. Genomic DNA was extracted from the iPSCs, and the primers were designed to amplify the eight exons of F IX. Next, the point mutation in those iPSCs was genetically corrected using CRISPR/Cas9 technology in the presence of a 129-nucleotide homologous repair template that contained two synonymous mutations. Then, top 8 potential off-target sites were subsequently analyzed using Sanger sequencing. Finally, the corrected clones were differentiated into hepatocyte-like cells, and the secretion of F IX was validated by immunocytochemistry and ELISA assay.Results The cell line bore a missense mutation in the 6th coding exon （c.676 C〉T） of F IX gene. Correction of the point mutation was achieved via CRISPR/Cas9 technology in situ with a high efficacy at about 22% （10/45） and no off-target effects detected in the corrected iPSC clones. F IX secretion, which was further visualized by immunocytochemistry and quantified by ELISA in vitro, reached about 6 ng/ml on day 21 of differentiation procedure. Conclusions Mutations in human disease-specific iPSCs could be precisely corrected by CRISPR/Cas9 technology, and corrected cells still maintained hepatic differentiation capability. Our findings might throw a light on iPSC-based personalized therapies in the clinical application, especially for hemophilia B.

Transarterial chemoembolization combined with recombinant human adenovirus type 5 H101 prolongs overall survival of patients with intermediate to advanced hepatocellular carcinoma: a prognostic nomogram study

Background: Patients with intermediate to advanced hepatocellular carcinoma(HCC) are most commonly treated with transarterial chemoembolization(TACE). Previous studies showed that TACE combined with recombinant human adenovirus type 5(H101) may provide a clinical survival benefit. In the present study, we aimed to determine the survival benefit of TACE with or without H101 for patients with intermediate to advanced HCC and to develop an e ective nomogram for predicting individual survival outcomes of these patients.Methods: We retrospectively collected data from 590 patients with intermediate to advanced HCC who were treated at Sun Yat?sen University Cancer Center between January 2007 and July 2015. After propensity score matching, 238 patients who received TACE with H101(TACE with H101 group) and 238 patients who received TACE without H101(TACE group) were analyzed. Overall survival(OS) was evaluated using the Kaplan–Meier method; the nomogram was developed based on Cox regression analysis. Discrimination and calibration were measured using the concordance index(c?index) and calibration plots.Results: Clinical and radiologic features were similar between the two groups. OS rates were significantly lower in the TACE group than in the TACE with H101 group(1?year OS rate, 53.8% vs. 61.3%; 2?year OS rate, 33.4% vs. 44.2%; 3?year OS rate, 22.4% vs. 40.5%; all P < 0.05). Multivariate Cox regression analysis for the entire cohort showed that alpha?fetoprotein level, alkaline phosphatase level, tumor size, metastasis, vascular invasion, and TACE with or without H101 were independent factors for OS, all of which were included in the nomogram. Calibration curves showed good agreement between nomogram?predicted survival and observed survival. The c?index of the nomogram for predict?ing OS was 0.716(95% confidence interval 0.686–0.746).Conclusions: TACE plus H101 extends the survival of patients with intermediate to advanced HCC. Our proposed nomogram provides individual survival prediction and stratification for patients with intermediate to advanced HCC who receive TACE with or without H101.

Antitumor activity of an hTERT promoter-regulated tumor-selective oncolytic adenovirus in human hepatocellular carcinoma

AIM: To construct a tumor-selective replication-competent adenovirus (RCAd), SG300, using a modified promoter of human telomerase reverse transcriptase (hTERT). METHODS: The antitumor efficacy of SG300 in hepatocellular carcinoma was assessed in vitro and in vivo. In vitro cell viability by MTT assay was used to assess the tumor-selective oncolysis and safety features of SG300, and in vivo antitumor activity of SG300 was assessed in established hepatocellular carcinoma models in nude mice. RESULTS: SG300 could lyse hepatocellular carcinoma cells at a low multiplicity of infection (MOI), but could not affect growth of normal cells even at a high MOI. Both in Hep3B and SMMC-7721 xenograft models of hepatocellular carcinoma, SG300 had an obvious antitumor effect, resulting in a decrease in tumor volume. Its selective oncolysis to tumor cells and safety to normal cells was also superior to that of ONYX-015. Pathological examination of tumor specimens showed that SG300 replicated selectively in cancer cells and resulted in apoptosis and necrosis of cancer cells. CONCLUSION: hTERT promoter-regulated replicativeadenovirus SG300 has a better cancer-selective replication-competent ability, and can specifically kill a wide range of cancer cells with positive telomerase activity, and thus has better potential for targeting therapy of hepatocellular carcinoma.

Detection of CYP2E1,a Genetic Biomarker of Susceptibility to Benzene Metabolism Toxicity in Immortal Human Lymphocytes Derived from the Han Chinese Population

Objective Cytochrome P450 2E1 （CYP2E1） is an important metabolizing enzyme involved in oxidative stress responses to benzene, a chemical associated with bone marrow toxicity and leukemia, We aimed to identify the CYP2E1 genetic biomarkers of susceptibility to benzene toxicity in support of environmental and occupational exposure prevention, and to test whether a model using immortal human lymphocytes might be an efficient tool for detecting genetic biomarkers. Methods Immortalized human lymphocyte cell lines with independent genotypes on four CYP2E1 SNP sites were induced with 0.01% phenol, a metabolite of benzene. CYP2E1 gene function was evaluated by mRNA expression and enzyme activity. DNA damage was measured by Single-Cell Gel Electrophoresis （SCGE）. Results Among the four SNPs, cells with rs2070673TT and rs2030920CC showed higher levels of ~YP2E1 transcription and enzymatic activity than the other genotypes in the same SNP site. Cells with higher gene expression genotypes also showed higher comet rates compared with lower gene expression genotypes. Conclusion These results suggest that CYP2E1 rs2070673 and rs2030920 might be the genetic biomarkers of susceptibility to benzene toxicity and that the immortalized human lymphocytes model might be an efficient tool for the detection of genetic biomarkers of susceptibility to chemicals.

Genetics of inflammatory bowel disease: The role of the HLA complex

The human leucocyte antigen （HLA） complex on chromosome 6p21.3 is the most extensively studied genetic region in Inflammatory bowel disease （IBD）. Consistent evidence of linkage to IBD3 （6p21.1-23）, an area which encompasses the HLA complex, has been demonstrated for both Crohn＇s disease and ulcerative colitis, and a number of replicated associations with disease susceptibility and phenotype have recently emerged. However, despite these efforts the HLA susceptibility gene （s） for IBD remain elusive, a consequence of strong linkage disequilibrium, extensive polymorphism and high gene density across this region. This article reviews current knowledge of the role of HLA complex genes in IBD susceptibility and phenotype, and discusses the factors currently limiting the translation of this knowledge to clinical practice.

Genetic diversity for grain Zn concentration in finger millet genotypes:Potential for improving human Zn nutrition

Nearly half of the world population suffers from micronutrient malnutrition,particularly Zn deficiency.It is important to understand genetic variation for uptake and translocation behaviors of Zn in relevant crop species to increase Zn concentration in edible parts.In the present study,genetic variation in grain Zn concentration of 319 finger millet genotypes was assessed.Large genetic variation was found among the genotypes,with concentrations ranging from 10 to 86 μg g^(-1)grain.Uptake and translocation studies with Zn/^(65) Zn application in 12 selected low-Zn genotypes showed wide variation in root uptake and shoot translocation,with genotypes GEC331 and GEC164 showing greater uptake and translocation.Genotypes GEC164 and GEC543 showed increased grain Zn concentration.Genotypes GEC331 and GEC164 also showed improved yield under Zn treatment.Appreciable variation in grain Zn concentration among finger millet genotypes found in this study offers opportunities to improve Zn nutrition through breeding.

Optimization of Biodynamic Seated Human Models Using Genetic Algorithms

Many biodynamic models have been derived using trial and error curve-fitting technique, such that the error between the computed and measured biodynamic response functions is minimum. This study developed a biomechanical model of the human body in a sitting posture without backrest for evaluating the vibration transmissibility and dynamic response to vertical vibration direction. In describing the human body motion, a three biomechanical models are discussed (two models are 4-DOF and one model 7-DOF). Optimization software based on stochastic techniques search methods, Genetic Algorithms (GAs), is employed to determine the human model parameters imposing some limit constraints on the model parameters. In addition, an objective function is formulated comprising the sum of errors between the computed and actual values (experimental data). The studied functions are the driving-point mechanical impedance, apparent mass and seat- to-head transmissibility functions. The optimization process increased the average goodness of fit and the results of studied functions became much closer to the target values (Experimental data). From the optimized model, the resonant frequencies of the driver parts computed on the basis of biodynamic response functions are found to be within close bounds to that expected for the human body.

Adenovirus-mediated expression of pig α(1,3) galactosyltransferase reconstructs Gal α(1,3) Gal epitope on the surface of human tumor cells

Gal alpha(1, 3) Gal (gal epitope) is a carbohydrate epitope and synthesized in large amount by alpha(1, 3) galactosyltransferase [alpha(1, 3) GT] enzyme on the cells of lower mammalian animals such as pigs and mice. Human has no gal epitope due to the inactivation of alpha(1, 3) GT gene but produces a large amount of antibodies (anti-Gal) which recognize Gal alpha(1, 3) Gal structures specifically. In this study, a replication-deficient recombinant adenoviral vector Ad5sGT containing pig alpha(1, 3) GT cDNA was constructed and characterized. Adenoviral vector-mediated transfer of pig alpha(1, 3) GT gene into human tumor cells such as malignant melanoma A375, stomach cancer SGC-7901, and lung cancer SPC-A-1 was reported for the first time. Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis, although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction. Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I (UEA I) lectins, Vicia villosa agglutinin (VVA), Arachis hypogaea agglutinin (PNA), and Glycine max agglutinin (SBA) to different degrees. In addition, no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

ADV36 adipogenic adenovirus in human liver disease

obesity and liver steatosis are usually described as related diseases.obesity is regarded as exclusive consequence of an imbalance between food intake and physical exercise,modulated by endocrine and genetic factors.Non-alcoholic fatty liver disease(NAFLD),is a condition whose natural history is related to,but not completely explained by over-nutrition,obesity and insulin resistance.There is evidence that environmental infections,and notably adipogenic adenoviruses(ADV)infections in humans,are associated not only with obesity,which is sufficiently established,but also with allied conditions,such as fatty liver.In order to elucidate the role,if any,of previous ADV36 infection in humans,we investigated association of ADV36-ADV37 seropositivity with obesity and fatty liver in humans.Moreover,the possibility that lifestyle-nutritional intervention in patients with NAFLD and different ADV36 seropositive status,achieves different clinical outcomes on ultrasound bright liver imaging,insulin resistance and obesity was challenged.ADV36 seropositive patientshave a more consistent decrease in insulin resistance,fatty liver severity and body weight in comparison with ADV36 seronegative patients,indicating a greater responsiveness to nutritional intervention.These effects were not dependent on a greater pre-interventional body weight and older age.These results imply that no obvious disadvantage-and,seemingly,that some benefit-is linked to ADV36 seropositivity,at least in NAFLD.ADV36 previous infection can boost weight loss and recovery of insulin sensitivity under interventional treatment.

Genetically modified human umbilical cord blood cells as a promising strategy for treatment of spinal cord injury

Spinal cord injury （SCI） continues to be a pressing health and social problem. The injury leads to neuronal and glial cell death accompanied by degeneration of nerve fibers. There are currently no particularly effective treatments. SCI causes profound disabil- ity of people affected and has attracted increased attention in the international field of neuroregeneration. For the past two decades, much hope has been placed in cell therapies for the restoration of both structure and function of the injured spinal cord. Embryonic and neural stem cells, olfactory ensheathing cells, microglia-like cells, Schwann cells, mesenchymal stem cells.

Advances in homology directed genetic engineering of human pluripotent and adult stem cells

The ability to introduce precise genomic modifications in human cells has profound implications for both basic and applied research in stem cells, ranging from identification of genes regulating stem cell self-renewal and multilineage differentiation to therapeutic gene correction and creation of in vitro models of human diseases. However, the overall efficiency of this process is challenged by several factors including inefficient gene delivery into stem cells and low rates of homology directed site-specific targeting. Recent studies report the development of novel techniques to improve gene targeting efficiencies in human stem cells; these methods include molecular engineering of viral vectors to efficiently deliver episomal genetic sequences that can participate in homology directed targeting, as well as the design of synthetic proteins that can introduce double-stranded breaks in DNA to initiate such recombination events. This review focuses on the potential of these new technologies to precisely alter the human stem cell genome and also highlights the possibilities offered by the combination of these complementary strategies.

Genetic diversity of Mansonia altissima A. Chev. under different regimes of human impact in the Akure Forest Reserve,Nigeria

Mansonia altissima is an important West African timber tree species. For the purpose of examining the effect of human impact on its genetic diversity, genetic diversity and spatial genetic structure of the species under different regimes of human impact were investigated in the Akure Forest Reserve, Nigeria, using 504 amplified fragment length polymorphism （AFLP） markers. The results indicate a very low genetic diversity in M. altissima within the forest reserve （He = 0.045; PPL = 16.75%; Br = 1.162）. The highest genetic diversity was observed in the primary forest （H e= 0.062; PPL - 21.00%; Br = 1.204）, with the lowest genetic diversity in the isolated forest patch （He = 0.032; PPL = 9.00%; B r= 1.089）. A significant and pronounced spatial genetic structure was found in the logged forest and in the isolated forest patch. In contrast, the primary forest exhibited very weak spatial genetic structuring. As expected, no spatial genetic structure was found in the planted stands of M. altissima. From a conservation point of view, our results suggest that genetic diversity ofM. altissima is at risk in the forest reserve. The scale of human impact in the study area could pose a serious threat to the maintenance of genetic diversity of the species. These results would offer practical applications in the conservation of other tropical tree species.

CONSTRUCTION OF A RECOMBINANT ADENOVIRUS VECTOR OF HUMAN PAPILLOMAVIRUS TYPE 16 L1_E7C

Objective: Human papillomaviruses are closely associated with human cervical cancer, especially HPV types 16 and 18. At present, HPV can not be produced in large quantity; it also has tumorgenicity and these properties of HPV have seriously hampered the development of HPV vaccine. HPV type 16 LI proteins can assembled into virus-like particles (VLP), which are morphologically identical to the nature virion. In order to develop the recombinant adenovirus vectors of HPV, we constructed a recombinant adenovirus shuttle plasmid pCA14 L1-E7C. Methods: Human papillomavirus type 16 LI open reading frame without terminator codon (TAA) (5559–7152) and E7C (682–855) were amplified using PCR. The L1 and E7C fragments were inserted into pGEM-T easy vectors by T-A strategy, named pTAL1 and pTAE7C. pTAL1 was cut with Hind III and BgIII, the pTAE7C with BamHI and ClaI. The L1 DNA fragment, E7C and pBluesscript SK were ligated together using T4 DNA ligase. pBSL1-E7C and pBSL1-E7C was digested with Hind III and Xhol. The L1-E7C fragment was inserted into adenovirus shuttle plasmids pCA14, named pCA14L1-E7C. DNA sequence results indicated that The L1-E7C DNA fragment can encode the HPV16L1-E7 fusion protein correctly. Results: The L1 and E7C DNA fragments were amplified by PCR and recombinant plasmid pTAL1, pTAE7C, pBSL1-E7C and pCA14L1-E7C were constructed correctly. The pCA14L1-E7C can be used in the further research work, cotransfected the 293 cell with the parent adenovirus pBHG10. Conclusion: Our results indicated that we have constructed a HPV16L1-E7 fusion DNA fragments and the adenovirus shuttle plasmids pCAL1-E7C for the further research.

Benefits of Genetic Engineering to the Environment and Human Health

The purpose of this essay is to argue that the genetic engineering may bring about benefits to human health and the environment.By means of research of secondary source collection,relevant evidence is selected,evaluated and organized into three main parts:improving agricultural environment,providing effective medical therapy and supplying safe and nutrition food to human body.In order to explain the benefits that created by genetic engineering technologies,examples based on opinions of experts and results of experts' experiments are used.The research results strongly suggest that the genetic engineering has positive effects on environment and mankind.Base on those finds,the argument is justified that genetic engineering is certainly beneficial to the environment and human health.In the future,more attention and researches should be focus on the genetic engineering with the purpose of benefiting human beings and the natural worlds.

Human a Type Genetic Engineering Interference Essence Injection

The highest-level interference essence against virus and turnour genetic engineering medicine is a new type created in the 1980s. Compared with chemical medicines, the interference essence has a special effect in the treatment of viruses and tumours. The human a, type genetic engineering interference essense is prepared by the Institute of Viruses of the Chinese Academy of Preventive Medical Sciences, the Shanghai Vaccine

Embryonic, genetic and clinical outcomes of fresh versus vitrified oocyte: A retrospective cohort study

Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam.The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified.In the last oocyte retrieval,all patient’s oocytes including both frozen and fresh would be fertilized.The outcomes included the rates of oocyte survival,cleavage embryo,blastocyst,ploidy status,pregnancy,biochemical pregnancy and clinical pregnancy.Results:The oocyte survival rate after thawing was 96.5%.No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate(78.1%vs.75.5%,P=0.461),usable cleavage embryo rate(86.9%vs.87.2%,P=0.916)but usable blastocyst rate was found higher statistically in the frozen oocyte group(44.4%vs.54.0%,P=0.049).The percentages of euploid,aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences(33.8%vs.31.6%,P=0.682;51.0%vs.54.2%,P=0.569;15.2%vs.12.4%,P=0.787;respectively).The rates of pregnancy,biochemical pregnancy and clinical pregnancy had no statistical difference(68.8%vs.64.8%,P=0.764;12.5%vs.3.6%,P=0.258;37.5%vs.46.4%,P=0.565).17 Mature oocytes are the minimum to have at least one euploid embryo.Conclusions:Oocyte vitrification does not affect embryonic,genetic and clinical results.The number of mature oocytes should be considered for fertilization in some cases.