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Dentritic cells (DCs) dysfunction has been verified detrimental for sepsis and B and T lymphocyte attenuator (BTLA) is an immune-regulatory receptor shown to be associated with DCs dysfunction. However, the role of BTLA expression in myeloid DCs (mDCs) in neonatal sepsis is unknown. In the current study, we found BTLA-expressing mDCs were elevated in neonates with sepsis and the BTLA expression level in mDCs was positively correlated to the severity of sepsis. The presence of BTLA negatively regulated the phagocytosis capacity and bactericidal ability of mDCs as well as the maturation markers expression of mDCs. Our data also showed BTLA+mDCs shifted into an anti-inflammatory phenotype with decreased expression of IL-6, TNF-α and IL-12, but increased IL-10. in addition, we found BTLA expression indeedly altered the mDCs allo-stimulatory capacity. Therefore, BTLA expression in mDCs could be a useful predictive marker for neonatal sepsis and targeting BTLA expression in mDCs may be a new therapeutic strategy.

Mitten crab (Eriocheir sensu stricto) is an indigenous and economically important species in East Asia. The genetic diversity and population structure of four wild native Eriocheir s. s. populations, from Yangtze (YZ), Oujiang (OJ), Minjiang (MJ), and Nanliujiang (NLJ) River basin, were studied. Genetic diversity of the four populations was evaluated by eight microsatellite and mitochondrial cytochrome oxidase subunits I (COI) markers. The polymorphism information content (PIC) values ranged from 0.7827 to 0.8580 and Shannon Wiener index (I) values varied from 2.0722 to 2.4088 respectively, according to microsatellite markers analysis. The haplotype diversity (h) values ranged from 0.52101 to 0.87097, while the values of nucleotide diversity (π) varied from 0.00139 to 0.02796 based on mitochondrial COI gene sequence. Four wild Eriocheir s. s. populations all showed high genetic diversity. As microsatellite analysis results, pair-wise FST values of YZ, OJ, MJ and NLJ populations were 0.1347, 0.1103 and 0.0943 respectively, which indicated that they were in medium genetic differentiation (0.05

Four previously undescribed compounds, including three glucosyloxybenzyl 2-isobutylmalates (1-3), one phenolic glycoside (4), along with ten known compounds were isolated from the flowers of Bletilla striata. The structures and absolute configurations of the undescribed compounds were elucidated on the basis of HR-ESIMS, NMR spectroscopy, optical rotation value, and acid hydrolysis experiment. Cytotoxicity of the isolated compounds against A549, HCT-116, and SW1990 cells and protective effects of t-BHP-induced L02 cytotoxic were assayed. The antioxidant activities of the isolated compounds were also evaluated.

Luteinizing hormone releasing hormone analog (LHRHa, des Gly10, [D-Ala6] ethylamide) is routinely applied for induced spawning of fish. Simulated gastric acid and pepsin were used to mimic human digestion in vitro to determine the stability of LHRHa upon possible ingestion. In vitro cleavage of LHRHa was quantified using UPLC-MS, ultra-performance liquid chromatography-mass spectrometry. LHRHa was broken down mainly into two fragments from Glp-His-Trp-Ser-Tyr-D-Ala-Leu-Arg-Pro-NHEt (LHRHa) to Trp-Ser-Tyr-D-Ala-Leu-Arg-Pro-NHEt (fragment 1) and Ser-Tyr-D-Ala-Leu-Arg-Pro-NHEt (fragment 2). By 24h of incubation, LHRHa was completely digested or barely detectable if the starting material was at 250 ng/μl. If the starting concentration was at 5 ng/μl, LHRHa was completely digested by 5h of incubation, or earlier. In both extreme scenarios, the results indicated that LHRHa would be digested completely in the human stomach and would pose no risk in human food consumption. This study supports the assertion that LHRHa used as a spawning aid would not pose a human food safety risk.

Clonorchis sinensis (C. sinensis) infections remain the common public health problem in freshwater fish consumption areas. New effective prevention strategies are still the urgent challenges to control this kind of foodborne infectious disease. The biochemical importance and biological relevance render C. sinensis enolase (Csenolase) as a potential vaccine candidate. In the present study, we constructed Escherichia coli/Bacillus subtilis shuttle genetic engineering system and investigated the potential of Csenolase as an oral vaccine candidate for C. sinensis prevention in different immunization routes. Our results showed that, compared with control groups, both recombinant Csenolase protein and nucleic acid could induce a mixed IgG1/IgG2a immune response when administrated subcutaneously (P

Common carp, Cyprinus carpio, is one of the most widely grown aquaculture species. Four types of knock-down constructs designed to transgenically sterilize fish were electroporated into common carp embryos to knockdown or eliminate expression of primordial germ cell (PGC) proteins, nanos or dead end. Embryos from different developmental stages were collected and tested using quantitative real-time PCR. Relative levels of gene expression were compared among control embryos and embryos exposed to these constructs during development. Two proteins, nanos and dead end, were targeted, each specific for a set of constructs, and a third related, but non-targeted PGC protein, vasa, was assayed. The effectiveness of the constructs varied depending upon the promoter. The expression levels of these proteins naturally decreased during development. In some, but not all cases, the expression of targeted mRNA was knocked down to minimal levels or degradation was accelerated. Up-regulation also was observed for dead end, possibly due to interferon responses. In other cases, the sterilization constructs had little effect on the targeted gene, but strongly down-regulated one or two of the alternative PGC genes examined in this study. At 19 months of age, common carp males exposed to these constructs had reduced rates of sexual maturity; however, reduction of female rates of sexual maturity was not conclusively shown. These constructs appear promising for transgenic sterilization of common carp.

We describe and compare normal eye, unilateral microphthalmia (small eye), and unilateral anophthalmia (eye incompletely developed) in young (5–7 month-old) common carp, Cyprinus carpio , (Cypriniformes: Cyprinidae). Five of 1058 (0.47%) common carp had grossly-observed eye deformities that we characterized using osteology and histopathology: 2 (81 and 98 mm fork length) had unilateral microphthalmia and 3 (103, 123, and 137 mm in fork length) had unilateral anophthalmia. Grossly, the eye of the microphthalmic common carp was opaque and slightly protruding from a reduced orbit; whereas, only a slight integumental concavity was observed on the deformed side of the anophthalmic common carp. Osteologically, microphthalmic and anophthalmic common carp had an irregular orbit with orbital bones that were laterally-expanded and that formed a discontinuous, uneven orbital perimeter compared to the narrow orbital bones forming a nearly circular orbit in normal common carp. Histologically, microphthalmia manifested as an eye having a thickened cornea with a retina and attached optic nerve but no lens (aphakia); whereas, anophthalmia manifested as lacking a globe, cornea, lens, or retina but having a nerve fascicle and capillaries representing probable optic nerve and its associated choroid. No histological sign of infection was observed in the ocular region of any common carp studied. The etiological agent associated with microphthalmia and anophthalmia is indeterminate, and the hypothesis that electroporation caused the ocular deformity was not tested herein. We suspect that the microphthalmic eye could detect light but not form a focused image. A list of putatively similar abnormalities in other fishes as well as anisophthalmia (incomplete eye development) and symmetrical cyclopia (medially fused eye) in larval and juvenile fishes is provided. This is the first published report providing osteological or histopathological details of microphthalmia or anophthalmia in a non-embryonic or non-larval fish.

25,27-Dipropoxy- p - tert -butylcalix[4]crown-9 cone conformer 5 was readily synthesized via an intramolecular cyclization strategy in good yields. The structures of all the new compounds involved were confirmed by NMR, ESI-MS and elemental analyses. All of them were proved to be in the cone conformation.

Brain levels of adenosine are elevated during hypoxia. Through effects on adenosine receptors (A(1), A(2A), A(2B) and A(3)) on astrocytes, adenosine can influence functions such as glutamate uptake, reactive gliosis, swelling, as well as release of neurotrophic and neurotoxic factors having an impact on the outcome of metabolic stress. We have studied the roles of these receptors in astrocytes by evaluating their susceptibility to damage induced by oxygen deprivation or exposure to the hypoxia mimic cobalt chloride (CoCl(2)). Hypoxia caused ATP breakdown and purine release, whereas CoCl(2) (0.8 mM) mainly reduced ATP by causing cell death in human D384 astrocytoma cells. Further experiments were conducted in primary astrocytes prepared from specific adenosine receptor knock-out (KO) and wild type (WT) mice. In WT cells purine release following CoCl(2) exposure was mainly due to nucleotide release, whereas hypoxia-induced intracellular ATP breakdown followed by nucleoside efflux. N-ethylcarboxamidoadenosine (NECA), an unselective adenosine receptor agonist, protected from cell death following hypoxia. Cytotoxicity was more pronounced in A(1)R KO astrocytes and tended to be higher in WT cells in the presence of the A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). Genetic deletion of A(2A) receptor resulted in less prominent effects. A(3)R KO glial cells were more affected by hypoxia than WT cells. Accordingly, the A(3) receptor agonist 2-chloro-N(6)-(3-iodobenzyl)-N-methyl-5'-carbamoyladenosine (CL-IB-MECA) reduced ATP depletion caused by hypoxic conditions. It also reduced apoptosis in human astroglioma D384 cells after oxygen deprivation. In conclusion, the data point to a cytoprotective role of adenosine mediated by both A(1) and A(3) receptors in primary mouse astrocytes.

Enterovirus 71 (EV71) is an etiologic agent responsible for seasonal epidemics of hand-foot-and-mouth disease and causes outbreaks with significant mortality among young children. To develop the vaccine, we have produced and purified the EV71 virus-like particle (VLP) that resembles the authentic virus in appearance, capsid structure and protein composition. In this study, we further evaluated the potential of VLP as a vaccine by comparing the humoral and cellular immune responses elicited by the purified VLP, denatured VLP and heat-inactivated EV71 virus. After immunization of BALB/c mice, EV71 VLP induced potent and long-lasting humoral immune responses as evidenced by the high total IgG titer and neutralization titer. The splenocytes collected from the VLP-immunized mice exhibited significant cell proliferation and produced high levels of IFN-gamma, IL-2 and IL-4 after stimulation, indicating the induction of Th1 and Th2 immune responses by VLP immunization. More importantly, the VLP immunization of mother mice conferred protection (survival rate up to 89%) to neonatal mice against the lethal (1000 LD(50)) viral challenge. Compared with the VLP immunization, immunization with denatured VLP and heat-inactivated EV71 elicited lower neutralization titers and conferred less effective protection to newborn mice, although they induced comparable levels of total IgG and cellular immune responses. These data collectively indicate the importance of the preservation of VLP structure and implicate the potential of VLP as a vaccine to prevent EV71 infection.

BACKGROUND/AIMS : Host genetic factor and hepatic fibrosis may predispose to risk for hepatocellular carcinoma (HCC). This study aimed to assess the association between tumor necrosis factor (TNF) α polymorphism and hepatic fibrosis, and risk for HCC. METHODS : One hundred eight pairs of gender-matched and age-matched patients with HCC and unrelated healthy controls were genotyped for TNF308.2 and TNF238.2 alleles with polymerase chain reaction and direct sequencing. RESULTS : The frequency of TNF308.1/TNF308.2 genotype in cases was higher than that in controls [odds ratio (OR) = 4.37]. Multivariate analysis indicated that TNF308.2 allele (OR = 3.23), hepatitis B surface antigen (OR = 17.17), and antibodies to hepatitis C virus (OR = 45.52) were independent risk factors for HCC. Surrogate markers for significant fibrosis implied that cases with the TNF308.2 allele have more advanced liver fibrosis. Moreover, multivariate analysis indicated that cirrhosis with Child-Pugh grade C, low serum albumin, and low platelet count were independent risk factors for carrying the TNF308.2 allele. CONCLUSIONS : TNF308.2 allele carriage and chronic hepatitis B virus/hepatitis C virus infection are independent risk factors for HCC. Carriage of the TNF308.2 allele correlates with disease severity and hepatic fibrosis, which may contribute to a higher risk for HCC.

A serosurvey targeting Hepatitis B virus (HBV)-vaccinated children born between 1986 and 1998 was conducted in 2001 in remote Taiwanese villages where a 1993 serosurvey indicated high vaccine failure. The HBV S antigen (HBsAg) seropositive rate among vaccinees of 3-6-year-old children in 2001 was significantly lower than that of 1993 and was higher among children who had a delayed vaccination schedule and received the plasma-derived vaccine. Vaccine escape variants were more prevalent among recipients of recombinant HBV vaccine residing in Hualien. Our study highlights the importance of continued monitoring of vaccinees for incidence of HBV infection in order to refine future vaccination policy.

During the 2003 Severe Acute Respiratory Syndrome (SARS) outbreak, traditional intervention measures such as quarantine and border control were found to be useful in containing the outbreak. We used laboratory verified SARS case data and the detailed quarantine data in Taiwan, where over 150,000 people were quarantined during the 2003 outbreak, to formulate a mathematical model which incorporates Level A quarantine (of potentially exposed contacts of suspected SARS patients) and Level B quarantine (of travelers arriving at borders from SARS affected areas) implemented in Taiwan during the outbreak. We obtain the average case fatality ratio and the daily quarantine rate for the Taiwan outbreak. Model simulations is utilized to show that Level A quarantine prevented approximately 461 additional SARS cases and 62 additional deaths, while the effect of Level B quarantine was comparatively minor, yielding only around 5% reduction of cases and deaths. The combined impact of the two levels of quarantine had reduced the case number and deaths by almost a half. The results demonstrate how modeling can be useful in qualitative evaluation of the impact of traditional intervention measures for newly emerging infectious diseases outbreak when there is inadequate information on the characteristics and clinical features of the new disease—measures which could become particularly important with the looming threat of global flu pandemic possibly caused by a novel mutating flu strain, including that of avian variety.

Reaction of dibutyltin dichloride with 4-chlorobenzohydroxamic acid and 4-chlorobenzoic acid in 1:1:1 stoichiometry yields the new title compound di- n -butyl(4-chlorobenzoxy)(4-chlorobenzohydroxamato)tin(IV) 1, which had been characterized by IR and 1 H, 13 C, 119 Sn NMR. Single X-ray crystal structure analysis has been determined for 1, which crystallizes as the five-coordinate mixed-ligand dibutyltin(IV) complex containing both monodentate and bidentate ligands. The crystal structure of the compound reveals that the tin atom adopts distorted trigonal bipyramidal geometry with the Bu groups trans to each other. The C–Sn–C angle is 145.9(3)°, and the Sn–O bonds are 2.083(4) and 2.158(4) A in one ligand and 2.242(4) A in the other. An unusual feature of the structure is the presence of a kind of intermolecular hydrogen bonding which serves to stabilize the crystal structure.

The betA gene from Escherichia coli encoding choline dehydrogenase was transferred into elite maize inbred DH4866 via Agrobacterium -mediated transformation. Among five transgenic maize lines investigated, four lines, lines 2, 3, 4 and 5, accumulated significantly higher levels of glycinebetaine than wild type plants (non-transformants). At 10 and 15 °C, transgenic lines 2, 3, 4 and 5 exhibited higher germination capacity as determined by the germination speed and final germination percentage, and displayed less inhibition in seedling shoot growth rate than wild type plants. In chilling conditions, the chilling tolerance, expressed in cell membrane damage, degree of chilling injury, survival rate, and photosynthesis was enhanced in plants of transgenic lines 2, 3, 4 and 5 as compared with wild type plants. The increase of total soluble sugars and the increase of total free amino acids that was mostly due to accumulation of alanine, glutamic acid, glycine and serine might play an important role of osmoregulation under stress conditions.

Enterovirus 71 (EV71) is a neurotrophic virus that causes seasonal morbidity and mortality in children throughout the world with increasing frequency in recent years. Because of the lack of an effective antiviral agent, primary prevention, including the development of effective vaccines, is a top priority in terms of control strategies. Poliovirus vaccine technology, both live attenuated and inactivated, killed virus vaccines, can be adopted for use with EV71 because of their relatedness. In this study, we have characterized a laboratory-adapted EV71 strain, YN3-4a, which exhibits different characteristics from those of its parent isolate, neu, in having a rapid growth rate in Vero cells, a larger plaque size, and a lower LD50 in newborn mice. The YN3-4a can be produced at a high viral titer of up to 1010 tissue culture infective dose (TCID50) when grown in Vero cells, an approved substrate for virus vaccine production. Mouse antiserum raised against YN3-4a can neutralize a broad range of strains of EV71 isolated at different times from a variety of geographic regions. On passage in Vero cells, YN3-4a remained genetically and phenotypically stable. Many of the above-described features, such as high viral yield, strong immunogenicity, broad-based antigenic coverage, and passage stability, are desirable features in a prototype virus for the development of an inactivated viral vaccine.

For assessing the role of circulating immune complexes (CIC) in acute hepatitis A, IgM- and IgG-specific CIC were determined, by C1q and conglutinin (K) assays, in 205 patients with acute hepatitis A and 60 healthy controls. The concentration of each type of CIC in patients was higher than healthy controls (P= 0.0001). CIC was a common feature of acute hepatitis A with 95.6% of cases having at least one abnormal test result. The prevalence of abnormal IgM class CIC was significantly higher than IgG class CIC. There were significantly inverse correlations between levels of IgM class CIC and interval between onset of symptoms and patient presentation. The prevalence of abnormal IgM CIC was higher in patients with higher alanine aminotransferase (P= 0.001) and patients with jaundice (P= 0.0002). In conclusion, IgM class CIC is the predominant CIC in acute hepatitis A and correlated with disease activity. CIC may play a role in the pathogenesis of acute hepatitis A.

We analyzed data from death certificates for all infant deaths in Taiwan from 1981 to 1988. During this 8-year period, the crude infant mortality rate decreased from 8.9 to 5.3 per 1000 live births. Deaths due to infectious diseases, which constituted a major fraction of this mortality, also declined from 3.4 to 1.2 per 1000 live births. While infant deaths due to nearly all causes declined, deaths due to injury and accidents rose from 0.62 to 0.71 per 1000 live births, and the sudden infant death rate rose form 0.13 to 0.46. Notable geographic differences included a high death rate in the small islands off the coast and in the eastern mountainous counties (9.1–11.2/1000 live births); this rate was twice that in Taipei (4.5/1000 live births). In addition, the level of urbanisation was also an important determinant of death rate; urban areas had much lower rates than rural areas. The highest rate (15.4) was persistently observed in the rural areas where the aboriginal tribes reside. This differential rate between urban and rural areas was most prominent for the vaccine-preventable diseases; the aboriginal areas had rates that were 12.9 times those in the cities. Moreover, since neonatal deaths are severely under-reported in Taiwan, especially in less urbanised areas, our data presumably underestimate the urban-rural health differences. Our findings identify high risk areas for various causes of infant death, and indicate that more targeted intervention such as improving education and health care as well as environmental hygiene in some specific areas may be warranted.

Although the importance of diarrhea as a prime cause of morbidity and death in developing countries is well recognized, the disease burden in the United States has never been thoroughly examined. We have prepared national estimates of the annual number of cases of diarrhea in children less than 5 years of age and of the outcome, measured in terms of visits to a physician, hospitalizations, and deaths. The annual number of diarrheal episodes was estimated by reviewing longitudinal studies of childhood diarrhea conducted in the United States and extrapolating these data to the nation. Estimates of physician visits, hospitalizations, and deaths were prepared from a variety of national data sources. We estimate that 16.5 million children less than 5 years of age have between 21 and 37 million episodes of diarrhea annually. Of these, 2.1 to 3.7 million episodes lead to a physician visit, a total of 220,000 patients are hospitalized, and 325 to 425 children die. The major cost of diarrhea lies in the high numbers and cost of hospitalizations, because approximately 10.6% of hospitalizations in this age group are for diarrhea. Diarrheal deaths occur in relatively small numbers, are more common in the South and among black persons, are potentially avoidable, and could represent as much as 10% of the preventable postneonatal infant death in the United States. These estimates underscore the extensive burden of diarrheal illness in children in the United States and suggest that interventions to prevent disease or decrease its severity could be cost-effective.