Pathogenicity and landscape of differential gene expression in mice orally infected with clinical coxsackievirus A6 (CA6).

Hand, foot, and mouth disease (HFMD) is caused by more than 20 pathogenic enteroviruses belonging to the Picornaviridae family and Enterovirus genus. Since the introduction of the enterovirus-71 (EV71) vaccine in 2016, the number of HFMD cases caused by EV71 has decreased. However, cases of infections caused by other enteroviruses, such as coxsackievirus A6 (CA6) and coxsackievirus A10, have been increasing accordingly. In this study, we used a clinical isolate of CA6 to establish an intragastric infection mouse model using 7-day-old mice to mimic the natural transmission route, by which we investigated the differential gene expression profiles associated with virus infection and pathogenicity. After intragastric infection, mice exhibited hind limb paralysis symptoms and weight loss, similar to those reported for EV71 infection in mice. The skeletal muscle was identified as the main site of virus replication, with a peak viral load reaching 2.31 × 107 copies/mg at 5 dpi and increased infiltration of inflammatory cells. RNA sequencing analysis identified differentially expressed genes (DEGs) after CA6 infection. DEGs in the blood, muscle, brain, spleen, and thymus were predominantly enriched in immune system responses, including pathways such as Toll-like receptor signaling and PI3K-Akt signaling. Our study has unveiled the genes involved in the host immune response during CA6 infection, thereby enhancing our comprehension of the pathological mechanism of HFMD.IMPORTANCEThis study holds great significance for the field of hand, foot, and mouth disease (HFMD). It not only delves into the disease's etiology, transmission pathways, and severe complications but also establishes a novel mouse model that mimics the natural coxsackievirus A6 infection process, providing a pivotal platform to delve deeper into virus replication and pathogenic mechanisms. Additionally, utilizing RNA-seq technology, it unveils the dynamic gene expression changes during infection, offering valuable leads for identifying novel therapeutic drug targets. This research has the potential to enhance our understanding of HFMD, offering fresh perspectives for disease prevention and treatment and positively impacting children's health worldwide.

Host Restriction Factor A3G Inhibits the Replication of Enterovirus D68 by Competitively Binding the 5' Untranslated Region with PCBP1.

The host restriction factor APOBEC3G (A3G) inhibits an extensive variety of viruses, including retroviruses, DNA viruses, and RNA viruses. Our study shows that A3G inhibits enterovirus 71 (EV71) and coxsackievirus A16 (CA16) via competitively binding the 5' untranslated region (UTR) with the host protein poly(C)-binding protein 1 (PCBP1), which is required for the replication of multiple EVs. However, whether A3G inhibits other EVs in addition to EV71 and CA16 has not been investigated. Here, we demonstrate that A3G could inhibit the replication of EVD68, which requires PCBP1 for its replication, but not CA6, which does not require PCBP1 for replication. Further investigation revealed that the nucleic-acid-binding activity of A3G is required for EVD68 restriction, similar to the mechanism presented for EV71 restriction. Mechanistically, A3G competitively binds to the cloverleaf (1 to 123 nucleotides [nt]) and the stem-loop IV (234 to 446 nt) domains of the EVD68 5' UTR with PCBP1, thereby inhibiting the 5' UTR activity of EVD68; by contrast, A3G does not interact with CA6 5' UTR, resulting in no effect on CA6 replication. Moreover, the nonstructural protein 2C, encoded by EVD68, overcomes A3G suppression by inducing A3G degradation via the autophagy-lysosome pathway. Our findings revealed that A3G might have broad-spectrum antiviral activity against multiple EVs through this general mechanism, and they might provide important information for the development of an anti-EV strategy. IMPORTANCE As the two major pathogens causing hand, foot, and mouth disease (HFMD), enterovirus 71 (EV71) and coxsackievirus A16 (CA16) attract a lot of attention for the study of their pathogenesis, their involvement with cellular proteins, and so on. However, other EVs such as CA6 and EVD68 constantly occur sporadically or have spread worldwide in recent years. Therefore, more information related to these EVs is needed in order to develop a broad-spectrum anti-EV inhibitor. In this study, we first reveal that the protein poly(C)-binding protein 1 (PCBP1), involved in PV- and EV71 virus replication, is also required for the replication of EVD68, but not for the replication of CA6. Next, we found that the host-restriction factor A3G specifically inhibits the replication of EVD68, but not the replication of CA6, by competitively binding to the 5' UTR of EVD68 along with PCBP1. Our findings broaden knowledge related to EV replication and the interplay between EVs and host factors.

Two Patient Studies of a Companion Diagnostic Immuno-Positron Emission Tomography (PET) Tracer for Measuring Human CA6 Expression in Cancer for Antibody Drug Conjugate (ADC) Therapy.

An antigen binding fragment (BFab) derived from a tumor-associated mucin 1-sialoglycotope antigen (CA6) targeting antibody (huDS6) was engineered. We synthesized a companion diagnostic positron emission tomography (PET) tracer by radiolabeling BFab with [64Cu] to measure CA6 expression on cancer tissues prior to anti-human CA6 (huDS6-DM4 antibody-drug conjugate) therapy for ovarian and breast cancer patients. After chemotherapy, the ovarian patient received PET scan with 18F-2-fluoro-2-deoxyglucose ([18F]FDG: 10 mCi), followed by [64Cu]-DOTA-BFab ([64Cu]BFab; 5.5 mCi) 1 week later for PET scanning of CA6 expression and subsequent surgery. The breast cancer patient was treated with chemotherapy before primary tumor resection and subsequent [18F]FDG-PET scan. 4 weeks later the patient received of [64Cu]BFab (11.7 mCi) for CA6 PET scan. Whole body [18F]FDG-PET of the breast cancer patient indicated FDG-avid tumor metastases to the liver, bilateral hila and thoracic spine, but no uptake was observed for the ovarian patient. Each patient was also imaged by PET/CT with [64Cu]BFab at 1 and 24 hours after tracer administration. The [64Cu]BFab tracer was well tolerated by both patients without adverse effects, and no significant tracer uptake was observed in both patients. Immunohistochemistry (IHC) data indicated CA6 expressions were weak to intermediate and matched with the [64Cu]BFab-PET signals.

Single Nucleotide Polymorphism in the Aetiology of Caries: Systematic Literature Review.

Recent progress in the field of molecular biology and techniques of DNA sequence analysis allowed determining the meaning of hereditary factors of many common human diseases. Studies of genetic mechanisms in the aetiology of caries encompass, primarily, 4 main groups of genes responsible for (1) the development of enamel, (2) formation and composition of saliva, (3) immunological responses, and (4) carbohydrate metabolism. The aim of this study was to present current knowledge about the influence of single nucleotide polymorphism (SNP) genetic variants on the occurrence of dental caries. PubMed/Medline, Embase, and Cochrane Library databases were searched for papers on the influence of genetic factors connected with SNP on the occurrence of dental caries in children, teenagers, and adults. Thirty original papers written in English were included in this review. Study groups ranged from 30 to 13,000 subjects. SNPs were observed in 30 genes. Results of the majority of studies confirm the participation of hereditary factors in the aetiology of caries. Three genes, AMELX, AQP5, and ESRRB, have the most promising evidence based on multiple replications and data, supporting a role of these genes in caries. The review of the literature proves that SNP is linked with the aetiology of dental caries.

TAS2R38 and CA6 genetic polymorphisms, frequency of bitter food intake, and blood biomarkers among elderly woman.

OBJECTIVE: Taste sensitivity is one of the most important biological determinants of food choice. Three SNPs of the TAS2R38 gene (rs713598, rs1726866, and rs10246939) give rise to two common haplotypes: PAV and AVI. These haplotypes, as well as an SNP within the CA6 gene (rs2274333) that encodes carbonic anhydrase VI (CA6), correlate with bitterness perception. The extent of consumption of bitter food may influence some health outcomes. The aim of this study is thus to investigate the impact of the TAS2R38 and CA6 genetic polymorphisms on the choice of bitter food, BMI, blood lipoprotein, and glucose concentrations as well as systemic inflammation in elderly women. METHODS: The associations between the TAS2R38 diplotype, CA6 genotype, and the intake of bitter-tasting foods were studied in a group of 118 Polish women over 60 years of age. The intake of Brassica vegetables, grapefruit, and coffee was assessed using a food frequency questionnaire. Biochemical parameters were measured using the spectrophotometric method. Genotyping was performed using the high resolution melting method. RESULTS: We found a correlation between lipid profile, glucose and CRP levels, and frequency of bitter food intake. The AVI/AVI subjects drank coffee more frequently than did the PAV/PAV homozygotes, as did the A carriers of CA6 in comparison with the GG homozygotes. We also observed that simultaneous carriers of the PAV haplotype and A allele of TAS2R38 and CA6, respectively, choose white cabbage more frequent and had lower plasma levels of CRP and glucose than did AVI/AVI and GG homozygotes. CONCLUSIONS: In elderly women, the TAS2R38 and CA6 polymorphisms may affect the frequency of consumption of coffee and white cabbage, but not of other bitter-tasting foods.

Optical properties and energy transfer of Eu2+/Ce3+ co-activated Na3Ca6(PO4)5 phosphor.

Ce(3+) /Eu(2+) co-doped Na3 Ca6 (PO4 )5 phosphors were prepared using a combustion-assisted synthesis method. X-Ray powder diffraction (XRD) analysis confirmed the formation of a Na3 Ca6 (PO4 )5 crystal phase. Na3 Ca6 (PO4 )5 :Eu(2+) phosphors have an efficient bluish-green emission band that peaks at 489 nm, whereas Ce(3+) -doped Na3 Ca6 (PO4 )5 showed a bright emission band at 391 nm. Analysis of the experimental results suggests that enhancement of the Eu(2+) emission intensity in co-doped Na3 Ca6 (PO4 )5 :Eu(2+) ,Ce(3+) phosphors is due to a resonance-type energy transfer from Ce(3+) to Eu(2+) ions, which is predominantly governed by an exchange interaction mechanism. These results indicate that Ce(3+) /Eu(2+) co-doped Na3 Ca6 (PO4 )5 is potentially useful as a highly efficient, bluish-green emitting, UV-convertible phosphor for white-light-emitting diodes.

Gene Methylation Affects Salivary Levels of the Taste Buds' Trophic Factor, Gustin Protein.

The salivary protein, Gustin/carbonic anhydrase VI, has been described as a trophic factor responsible for the growth of taste buds. We found, in a genetically homogeneous population, that the polymorphism rs2274333 (A/G) of the Gustin gene is crucial for the full functionality of the protein and is associated with taste sensitivity. However, other studies have failed to find this evidence. Here, we verified if Gustin gene methylation can affect the salivary levels of the protein, also concerning the polymorphism rs2274333 and PROP bitter responsiveness. The Gustin gene methylation profiling and the quantification of the Gustin salivary levels were determined in sixty-six volunteers genotyped for the polymorphism rs2274333 (A/G) (Ser90Gly in the protein sequence). The fungiform papillae density was also determined. The results confirm our earlier observations by showing that AA genotypes had a greater density of fungiform taste papillae, whereas the GG genotypes showed a lower density. We also found variations in the protein levels in the three genotype groups and an inverse relationship between Gustin gene methylation and the salivary levels of the protein, mostly evident in AA and ST volunteers, i.e., in volunteers who would be carriers of the functional isoform of the protein. These findings could justify the conflicting data in the literature.

Investigating the Action Mechanism of Titanium in Alumina-Magnesia Castables by Adding Different Ti-Bearing Compounds.

To investigate the action mechanism of titanium, the effects of different Ti-bearing compounds, including CaTiO3, MgTiO3, and nano-TiO2, on the properties of alumina-magnesia castables were studied. By analyzing the phase compositions, microstructures, and physical and mechanical properties of the castables, it was demonstrated that an intermediate product, CaTiO3, was first generated. This was then consumed by solid-solution reactions, and titanium was involved in the liquid formation as the temperature increased. The solid-solution reaction of CA6 (CaAl12O19) was more prominent due to the incorporation of more titanium in the crystal lattice of CA6 instead of spinel (MgAl2O4). Moreover, the liquid formation was strongly promoted when more titanium accompanied the calcium, which finally accelerated the densification and improved the strengths of alumina-magnesia castables. On the whole, castables with CaTiO3 addition presented higher bulk density and excellent strength after the heat treatment. Besides, the castables with 2 wt.% CaTiO3 contents were estimated to possess greater thermal shock resistance.

Biochemical and Biophysical Characterization of Carbonic Anhydrase VI from Human Milk and Saliva.

Carbonic anhydrases (CA, EC 4.2.1.1) catalyze the hydration of carbon dioxide and take part in many essential physiological processes. In humans, 15 CAs are characterized, including the only secreted isoenzyme CA VI. CA VI has been linked to specific processes in the mouth, namely bitter taste perception, dental caries, and maintenance of enamel pellicle, and implicated in several immunity-related phenomena. However, little is known of the mechanisms of the above. In this study, we characterized human CA VI purified from saliva and milk with biophysical methods and measured their enzyme activities and acetazolamide inhibition. Size-exclusion chromatography showed peaks of salivary and milk CA VI corresponding to hexameric state or larger at pH 7.5. At pH 5.0 the hexamer peaks dominated. SDS- PAGE of milk CA VI protein treated with a bifunctional crosslinker further confirmed that a majority of CA VI is oligomers of similar sizes in solution. Mass spectrometry experiments confirmed that both of the two putative N-glycosylation sites, Asn67 and Asn256, are heterogeneously glycosylated. The attached glycans in milk CA VI were di- and triantennary complex-type glycans, carrying both a core fucose and 1 to 2 additional fucose units, whereas the glycans in salivary CA VI were smaller, seemingly degraded forms of core fucosylated complex- or hybrid-type glycans. Mass spectrometry also verified the predicted signal peptide cleavage site and the terminal residue, Gln 18, being in pyroglutamate form. Thorough characterization of CA VI paves way to better understanding of the biological function of the protein.

Salivary proteomic analysis in asymptomatic and symptomatic SARS-CoV-2 infection: Innate immunity, taste perception and FABP5 proteins make the difference.

BACKGROUND AND AIM: SARS-CoV-2 infection spawns from an asymptomatic condition to a fatal disease. Age, comorbidities, and several blood biomarkers are associated with infection outcome. We searched for biomarkers by untargeted and targeted proteomic analysis of saliva, a source of viral particles and host proteins. METHODS: Saliva samples from 19 asymptomatic and 16 symptomatic SARS-CoV-2 infected subjects, and 20 controls were analyzed by LC-MS/MS for untargeted peptidomic (flow through of 10 kDa filter) and proteomic (trypsin digestion of filter retained proteins) profiling. RESULTS: Peptides from 53 salivary proteins were identified. ADF was detected only in controls, while IL1RA only in infected subjects. PRPs, DSC2, FABP5, his-1, IL1RA, PRH1, STATH, SMR3B, ANXA1, MUC7, ACTN4, IGKV1-33 and TGM3 were significantly different between asymptomatic and symptomatic subjects. Retained proteins were 117, being 11 highly different between asymptomatic and symptomatic (fold change ≥2 or ≤-2). After validation by LC-MS/MS-SRM (selected reaction monitoring analysis), the most significant discriminant proteins at PCA were IL1RA, CYSTB, S100A8, S100A9, CA6, and FABP5. CONCLUSIONS: The differentially abundant proteins involved in innate immunity (S100 proteins), taste (CA6 and cystatins), and viral binding to the host (FABP5), appear to be of interest for use as potential biomarkers and drugs targets.

Coxsackievirus A6 Induces Necroptosis for Viral Production.

Hand, foot, and mouth disease (HFMD) is a febrile exanthematous disease with typical or atypical symptoms. Typical HFMD is usually caused by enterovirus 71 (EV71) or coxsackievirus A16, while atypical HFMD is usually caused by coxsackievirus A6 (CA6). In recent years, worldwide outbreaks of CA6-associated HFMD have dramatically increased, although the pathogenic mechanism of CA6 is still unclear. EV71 has been established to induce caspase-dependent apoptosis, but in this study, we demonstrate that CA6 infection promotes a distinct pathway of cell death that involves loss of cell membrane integrity. Necrostatin-1, an inhibitor of necroptosis, blocks the cell death induced by CA6 infection, but Z-DEVD-FMK, an inhibitor of caspase-3, has no effect on CA6-induced cell death. Furthermore, CA6 infection up-regulates the expression of the necroptosis signaling molecule RIPK3. Importantly, necrostatin-1 inhibits CA6 viral production, as assessed by its ability to inhibit levels of VP1 protein and genomic RNA and infectious particles. CA6-induced necroptosis is not dependent on the generation of reactive oxygen species; however, viral 3D protein can directly bind RIPK3, which is suggestive of a direct mechanism of necroptosis induction. Therefore, these results indicate that CA6 induces a mechanism of RIPK3-dependent necroptosis for viral production that is distinct from the mechanism of apoptosis induced by typical HFMD viruses.

Physicochemical, techno-functional, and antioxidant properties of a novel bacterial exopolysaccharide in cooked beef sausage.

This work investigates the effects of partial replacement of vitamin C (Vit C) with a purified exopolysaccharide (EPS-Ca6) produced by Lactobacillus sp. Ca6, on the antioxidant activities of cooked beef sausages during refrigerated storage. The physicochemical, techno-functional and viscosity properties of EPS-Ca6 were also studied. Functional properties of EPS-Ca6 were determined based on Water Holding Capacity (WHC), Oil Holding Capacity (OHC), emulsification activity, and foaming ability. EPS-Ca6 demonstrated excellent emulsifying and emulsion stabilizing properties. It was able to emulsify several food-grade oils and hydrophobic compounds, particularly corn oil and diesel with emulsification indexes of 90 and 100%, respectively at a concentration of 0.5%. The effect of EPS-Ca6 on oxidative processes in cooked beef sausages during storage up to 12days at 4°C was evaluated. The obtained results showed a high rate (p<0.05) of oxymyoglobin (OxyMb) and low lipid oxidation. Overall, our findings provided evidence that EPS-Ca6 could be used as a natural additive for maintaining storage stability of cooked beef sausages, and could replace synthetic polymer in several industrial applications.

Variations in the bitterness perception-related genes TAS2R38 and CA6 modify the risk for colorectal cancer in Koreans.

Bitterness perception is known to be an important factor in individuals' dietary behaviors and is also associated with the sensing of nutritious/noxious molecules for subsequent metabolic responses in multiple organs. Therefore, the genetic variation in bitterness sensing may be associated with diet-related diseases, including colorectal cancer (CRC). We investigated the influence of variations in the bitterness-sensing genes taste receptor type 2 member 38 (TAS2R38) and carbonic anhydrase 6 (CA6) on the consumption of food, tobacco and alcohol and the risk of CRC in Koreans. The study population consisted of 681 cases and 1361 controls, and their intake of vegetables, fruits, fiber, fat-food and sweets was analyzed. The genotypes for TAS2R38 A49P, V262A and I296V and CA6 rs2274333 A/G were assessed using the MassArray technique. Our findings suggested that the TAS2R38 diplotype, CA6 rs2274333 and their combined genotype had a negligible influence on dietary and alcohol intake. The combined TAS2R38-CA6 AVI/AVI-AA genotype was associated with higher tobacco consumption than the other genotypes in CRC cases only. However, the genetic variations were a significant risk factor for CRC. The TAS2R38 AVI/AVI diplotype and CA6 G allele were associated with a reduced risk of CRC. Moreover, when the combined genotypes of the subjects were analyzed, possessing both the variant diplotype/variant allele (AVI/AVI+G*) was associated with a greater reduction in the risk of CRC (adjusted OR = 0.49; 95%CI: 0.34-0.74). In summary, variations in the bitterness perception genes TAS2R38 and CA6 did not influence the examined food intake in Koreans. However, those genetic variants were a decisive modifying factor of CRC susceptibility.

Evaluation of dermal wound healing activity and in vitro antibacterial and antioxidant activities of a new exopolysaccharide produced by Lactobacillus sp.Ca6.

The present study aimed to evaluate the in vitro antibacterial and antioxidant activities and the in vivo wound healing performance of a noval exopolysaccharides (EPS-Ca6) produced by Lactobacillus sp.Ca6 strain. The results showed that EPS-Ca6 had a potential antioxidant activity determined through four different assays: DPPH scavenging activity, reducing power, ß-carotene bleaching by linoleic acid assay, and Metal chelating activities. It also exhibited significant antibacterial activity against pathogenic bacteria Salmonella enterica and Micrococcus luteus. The wound healing activity of the EPS-Ca6, using excision wound model in rats, showed that this novel EPS accelerated significantly wound healing activity as compared to the control group, and a total closure was achieved after 14days of wound induction. Furthermore, histological examination of biopsies showed fully re-epithelialized wound with a complete epidermal regeneration. Overall the finding indicates that the EPS-Ca6 might be useful as a wound healing agent in modern medicine.

Development of a molecular method for the rapid screening and identification of the three functionally relevant polymorphisms in the human TAS2R38 receptor gene in studies of sensitivity to the bitter taste of PROP.

The objective of this work was to develop a rapid screening method to identify the three single nucleotide polymorphisms (SNPs) in the TAS2R38 gene, with the aim of providing a significant contribution to studies designed to assess sensitivity to the bitter taste of 6-n-propylthiouracil (PROP). Specifically, the objective of this study was to characterize the TAS2R38 gene haplotypes in a group of 60 subjects with variable sensitivity to PROP and preliminarily genotyped for the rs2274333 allele (A/G) of carbonic anhydrase isoform VI gene (CA6). The molecular characterization of the TAS2R38 gene was conducted using the PCR-restriction fragment length polymorphism technique after creating artificial restriction sites upstream or downstream of the SNPs, as none of the three polymorphisms contributes to the formation of a restriction site for a specific endonuclease. The results indicate that the method described in this paper could be a valid and simple experimental strategy to identify genetic differences related to taste sensitivity to bitter taste, and could be applied as a nutrigenetics test in studies aimed at understanding people's eating behaviors.

Phylogenetic analysis of the major causative agents of hand, foot and mouth disease in Suzhou City, Jiangsu province, China, in 2012-2013.

Hand, foot and mouth disease (HFMD) is a serious public health problem that has emerged over the past several decades. Pathogen detection by the Chinese national HFMD surveillance system has focused mainly on enterovirus 71 (EV71) and coxsackievirus A16 (CA16). Therefore, epidemiological information regarding the other causative enteroviruses is limited. To identify the pandemic enterovirus in Suzhou, Jiangsu province, China, clinical samples from patients with HFMD were collected from 2012 to 2013 and analyzed. The results revealed that CA16 was the most dominant HFMD pathogen in 2012, whereas CA6 and CA10 were the dominant pathogens in 2013. Phylogenetic analysis revealed that the C4a sub-genogroup of EV71 and the B1a and B1b sub-genogroups of CA16 continued to evolve and circulate in Suzhou. The CA6 strains were assigned to six genotypes (A-F) and the CA10 strains were assigned to seven genotypes (A-G), with clear geographical and temporal distributions. All of the CA6 strains in Suzhou belonged to genogroup F, and there were several lineages circulating in Suzhou. All of the CA10 strains in Suzhou belonged to genogroup G, and they had the same genetic origin. Co-infections of EV71/CA16 and CA6/CA10 were found in the samples, and bootscan analysis of 5'-untranslated regions (UTRs) revealed that some CA16 strains in Suzhou had genetic recombination with EV71. This property might allow CA16 to alter its evolvability and circulating ability. This study underscores the need for surveillance of CA6 and CA10 in the Yangtze River Delta and East China.

The gustin (CA6) gene polymorphism, rs2274333 (A/G), is associated with fungiform papilla density, whereas PROP bitterness is mostly due to TAS2R38 in an ethnically-mixed population.

PROP responsiveness is associated with TAS2R38 haplotypes and fungiform papilla density. Recently, we showed that a polymorphism in the gene coding for the salivary trophic factor, gustin (CA6), affects PROP sensitivity by acting on cell growth and fungiform papillae maintenance, in a genetically homogeneous cohort. Since population homogeneity can lead to over estimation of gene effects, the primary aim of the present work was to confirm gustin's role in PROP bitterness intensity and fungiform papillae density in a genetically diverse population. Eighty subjects were genotyped for both genes by PCR techniques. PROP responsiveness was assessed by a filter paper method and fungiform papilla density was determined in each subject. As expected, PROP bitterness ratings were lower in individuals with the AVI/AVI diplotype of TAS2R38 than in individuals with PAV/PAV and PAV/AVI diplotypes. However, no differences in PROP bitterness among genotypes of the gustin gene, and no differences in the density of fungiform papillae related to TAS2R38 diplotype were found. In contrast, the density of fungiform papillae decreased as the number of minor (G) alleles at the gustin locus increased. In addition, the distribution of TAS2R38 genotypes within each gustin genotype group showed that the occurrence of recessive alleles at both loci was infrequent in the present sample compared to other populations. These findings confirm that papillae density is associated with gustin gene polymorphism, rs2274333 (A/G), in an ancestrally heterogeneous population, and suggest that variations in the frequency of allele combinations for these two genes could provide a salient explanation for discrepant findings for gustin gene effects across populations.

Molecular epidemiology of HFMD in mainland China and discussion on prevention and control measures / 中华微生物学和免疫学杂志

Objective To summarize the epidemiological data of hand, foot and mouth disease (HFMD) in China from 2013 to 2017 and to analyze its molecular epidemiological characteristics in order to provide scientific basis for improving prevention and control measures. Methods China National Knowledge In-frastructure,Wanfang Database of China,Chinese VIP Journal Net and Pubmed were used to search epidemio-logical data of HFMD published in recent years. National notification data and surveillance data of HFMD in ma-inland China were obtained from the National Health and Family Planning Commission of China and the World Health Organization. Basic statistic tools were used for data analysis. Results From 2013 to 2016, the inci-dence rates of HFMD were 134. 37/100 000, 203. 16/100 000, 145. 30/100 000, 176. 62/100 000 and 140.46/100 000,respectively. Enterovirus 71(EV71),coxsackievirus A16(CA16),CA6 and CA10 were the predominant pathogens causing HFMD in the first half of 2013-2017. CA6 was the main epidemic strain in most areas of China. EV71 remained the predominant pathogen causing severe HFMD, but CA6, CA16 and CA10 were also critical pathogens of concern. The predominant strains of enteroviruses varied with year and region. Conclusion Although the EV71 vaccine has been approved since 2016, HFMD has not been controlled com-pletely in China. It is badly in need of more comprehensive surveillance of other types of enteroviruses and HFMD polyvaccine to improve the prevention and control of HFMD.

Status analysis of prevailing strains of non-EV71 and non-CA16 enteroviruse / 重庆医学

Objective To investigatethe constituents of enteroviruse which can cause hand‐foot‐and‐mouth disease(HFMD) ,and prevent against the development of new prevailing strains and provide basis for prevention and control of HFMD in Luoyang city . Methods Reverse‐transcription polymerase chain reaction (RT‐PCR) was used to identify the type of RNA of stool specimen of HFMD cases in Luoyang ,to distinguish the subtype of strain ,9 enterovirus‐positive specimens which coxcackie virusA16(CA16)‐and enterovirus71(EV71)‐negative were selected randomly for 5′UTR sequencing .strain‐specific primers were used to detect posi‐tive samples in 75 CA16‐ and EV71‐negative but other enterovirus‐positive specimens by using real time RT‐PCR .Composition characteristics of CA6 were further analyzed .Results Primary screening was conducted with the results of 5′UTR sequencing of 9 commonent erovirus‐positive specimens and there were 5 CA6‐positive ,which were prevailing strains;there were 20 CA6‐positive of 75 commonenterovirus‐positive specimens ,the proportion of CA6‐positive in enterovirus‐positive specimens was 29 .76% (25/84) . Thirteen samples were collected from male and 12 from female ,2 cases resulted in severe symptoms ,5 from city and 20 from coun‐try .Conclusion The composition of commonent erovirus which results in HFMD was complex and the proportion of CA6 was higher ,which should be pay more attention .

The Relationship between Single Nucleotide Polymorphisms of the Carbonic Anhydrase 6 Gene andPhenylthiocarbamide Taste Sensitivity, Taste Disorder / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The phenylthiocarbamide (PTC) and 6-n-propylthiouracil (PROP) taste sensitivity varies among individuals. Recently, it is reported that PROP taste responsiveness is associated with carbonic anhydrase 6 (CA6) gene polymorphism. The CA6 gene, a zinc metalloprotein in human saliva, is affected in taste function and might be correlated with gustatory diversity. The aim of this study was to examine whether PTC taste sensitivity and taste disorder is associated with the CA6 gene polymorphism rs2274327 (C/T), rs2274328 (A/C), and rs2274333 (A/G). SUBJECTS AND METHOD: A total of 217 healthy normal subjects were recruited as controls, and 50 taste disorder patients were recruited as experimental group. The polymorphisms of CA6 gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis. All statistical analyses were calculated using the statistical package for the social science software. Haplotypes were estimated by Haploveiw and the PHASE programs. RESULTS: The CA6 gene polymorphisms showed association with taste disorder but not with PTC sensitivity (taster/nontaster). The number of control subjects carrying AA genotype of single nucleotide polymorphism rs2274328 (A/C) in the CA6 gene was higher than the number of the subjects with taste disorder (p=0.048). However, there was no association between controls and taste disorder subjects in the haplotype analysis. CONCLUSION: These data suggest that the CA6 gene polymorphism rs2274328 could affect taste function impairment in patients with taste disorder. This observation requires a further functional study of gustin protein to clarify the association of the CA6 gene polymorphisms with the taste disorder and sensitivity.