Two new ant species of the genus Leptogenys (Hymenoptera, Formicidae) from Hainan, China, with a key to the known Chinese species.

Two new species of ponerine ants from Hainan Province, China, Leptogenyshainanensissp. nov. and L.zhouisp. nov., are delineated and depicted based on the worker caste. Leptogenyshainanensissp. nov. belongs to the L.leleji species group, with mandibles elongate, slender and curved, lacking a distinct masticatory margin. On the other hand, L.zhouisp. nov. belongs to the L.crassicornis species group, distinguished by its square head, smooth body, mandibles with a dentate masticatory margin, and short antennae. A key to workers for the known species of Leptogenys in China are provided and a map is provided for the newly described species.

Association between phase angle and diabetic peripheral neuropathy in Type 2 diabetes patients.

BACKGROUND: Diabetic peripheral neuropathy (DPN), the very prevalent microvascular complication of type 2 diabetes mellitus (T2DM), severely threatens the quality of life of diabetic patients. Thus, prevention of DPN is extremely important for public health, and the identification of potential biomarkers may help with early prevention. Our work determined the association between phase angles (PhAs) and the risk of DPN in T2DM patients. MATERIALS AND METHODS: This cross-sectional study recruited 697 T2DM patients from the Third Affiliated Hospital of Soochow University. All patients were divided into the non-diabetic peripheral neuropathy group and diabetic peripheral neuropathy group, and the DPN group was further divided into low, medium, and high diabetic foot ulcer risk groups according to vibration perception threshold results. Using univariate and multivariate logistic regression analyses continuously, the relationship between PhAs (arms, legs, trunk, and whole body) and the risk of DPN were evaluated. Further analysis was conducted on different subgroups of the study population. RESULTS: After adjusting for the potential covariates, multivariate logistic regression analyses indicated that PhAs of the arms, legs, and whole body correlated inversely with the risk of DPN. However, the PhA of the trunk had no significant correlation with DPN. According to the stratified subgroup analysis, the negative association between PhA of the whole body and the risk of DPN remained significant in the sex and BMI group except for participants with BMI ≥ 28 kg/m². CONCLUSIONS: For the first time, PhAs were acknowledged to be independently associated with DPN. Further exploration is needed to explain the findings.

Association Between the Water Distribution in the Human Body and 25-Hydroxyvitamin D Among the Type 2 Diabetes Mellitus Population: A Possible Pathway Between Vitamin D and Diabetic Nephropathy.

Purpose: To explore the association between the water distribution in the human body and 25-hydroxyvitamin D among patients with type 2 diabetes mellitus (T2DM), and to analyze whether water distribution is a mediator between vitamin D and T2DM complications. Patients and Methods: In total, 533 T2DM inpatients were included from August 1, 2016 to April 1, 2023. Water distribution indicators, whether from the whole body, arms, trunk, or legs, were measured to calculate the association with vitamin D using linear regression analysis. Subgroups based on age, sex, body mass index, and macrovascular complications were established to clarify changes in the association between vitamin D and water distribution in different populations. Mediation analysis was applied to evaluate the correlation between vitamin D, water distribution, and T2DM complications. Results: There was a negative correlation between 25-hydroxyvitamin D and the ratio of extracellular water (ECW)/total body water (TBW) both in the whole body (P=0.045, ß=-0.008) and in certain parts. A U-shaped restricted cubic spline curve further presented an inflection point (approximately 23 ng/mL 25-hydroxyvitamin D) in the relationship between 25-hydroxyvitamin D and the ECW/TBW of the whole body. A negative correlation was observed between ECW/TBW and vitamin D in the obese subgroup (P=0.015, ß=-0.038). In the total effect of vitamin D on diabetic nephropathy (DN), the mediation effect of ECW/TBW accounted for 15.44%. Conclusion: A correlation between vitamin D and water distribution was observed, and a high ECW/TBW was one of the pathways through which low vitamin D levels might affect DN.

Prediction of the Risk of Bone Mineral Density Decrease in Type 2 Diabetes Mellitus Patients Based on Traditional Multivariate Logistic Regression and Machine Learning: A Preliminary Study.

Purpose: There remains a lack of a machine learning (ML) model incorporating body composition to assess the risk of bone mineral density (BMD) decreases in type 2 diabetes mellitus (T2DM) patients. We aimed to use ML algorithms and the traditional multivariate logistic regression to establish prediction models for BMD decreases in T2DM patients over 50 years of age, and compare the performance of the two methods. Patients and Methods: This cross-sectional study was conducted among 450 patients with T2DM from 1 August 2016 to 31 December 2022. The participants were divided into a normal BMD group and a decreased BMD group. Traditional multivariate logistic regression and six ML algorithms were selected to construct male and female models. Two nomograms were constructed to evaluate the risk of BMD decreases in the male and female T2DM patients, respectively. The ML models with the highest area under the curve (AUC) were compared with the traditional multivariate logistic regression models in terms of discriminant ability and clinical applicability. Results: The optimal ML model was the extreme gradient boost (XGBoost) model. The AUCs of the traditional multivariate logistic regression and the XGBoost models were 0.722 and 0.800 in the male testing dataset, respectively, and 0.876 and 0.880 in the female testing dataset, respectively. The decision curve analysis results suggested that using the XGBoost models to predict the risk of BMD decreases obtained more net benefits compared with the traditional models in both sexes. Conclusion: We preliminarily proved that the XGBoost models outperformed most other ML models in both sexes and achieved higher accuracy than traditional analyses. Due to the limited sample size in the study, it is necessary to validate our findings in larger prospective cohort studies.

Characterization and Phylogenetic Implication of Complete Mitochondrial Genome of the Medicinal Ant Formica sinae (Hymenoptera: Formicidae): Genomic Comparisons in Formicidae.

Ants (Hymenoptera: Formicidae) are among the largest and most widespread families of terrestrial insects and are valuable to medical and ecological investigations. The mitochondrial genome has been widely used as a reliable genetic marker for species identification and phylogenetic analyses. To further understand the mitogenome-level characteristics of the congeneric Formicidae species, the complete mitogenome of Formica sinae (Hymenoptera: Formicidae) was sequenced, annotated, and compared with other 48 Formicidae species. The results showed that gene composition, content, and codon usage were conserved. The complete mitochondrial genome of F. sinae was 17,432 bp, including 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and one control region located between rrnS and trnM, which was 1,256 bp long, the longest of all sequenced species. Gene rearrangement was not detected in Formica species (Hymenoptera: Formicidae). All PCGs of F. sinae were initiated with ATN codons and terminated with the TAA codon. The overall nucleotide composition of F. sinae was AT-biased (83.51%), being 80.58% in PCGs, 86.68% in tRNAs, 87.10% in rRNAs, and 88.70% in the control region. Phylogenetic analyses indicated that each subfamily formed a strongly monophyletic group. Furthermore, F. sinae clustered with Formica fusca (Hymenoptera: Formicidae) and Formica selysi (Hymenoptera: Formicidae). This work enhances the genetic data of Formicidae and contributes to our understanding of their phylogenic relationship, evolution, and utilization.

Characterization of the complete mitochondrial genome of a harvesting ant Messor structor (Hymenoptera: Formicidae: Myrmicinae).

Messor structor (Latreille, 1798) is a keystone ant species in the genus Messor (Formicidae: Myrmicinae). Here, we reported the complete mitochondrial genome of M. structor. The circular mitogenome of M. structor is 17628 bp including 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and a control region. The base composition was AT-biased (84.07%). Phylogenetic analysis suggests that it is closely related to Aphaenogaster famelica. The mitochondrial genome of M. structor will be a good source for understanding molecular evolutionary studies of this species and related ant species.

A new species of the ant Platythyreaclypeata species group (Hymenoptera, Formicidae, Ponerinae) from continental Asia.

The Platythyreaclypeata species group is reviewed and three species, including one new species, P.homasawini sp. nov., are recognized. This species group is distinguished from the P.parallela species group by the reddish-brown body, the elliptical shape of the propodeal spiracle, the elongate antennal scape, and the distinctly narrowed posteriad space between frontal carinae. Platythyreahomasawini sp. nov., from Thailand and China, is described based on the worker caste. The type series of the new species was collected on the forest floor from dead wood in an advanced stage of decomposition. A key to the Oriental species of the genus Platythyrea based on the worker caste is provided.

A multi-faceted comparative perspective on elevational beta-diversity: the patterns and their causes.

The observed patterns and underlying mechanisms of elevational beta-diversity have been explored intensively, but multi-dimensional comparative studies remain scarce. Herein, across distinct beta-diversity components, dimensions and species groups, we designed a multi-faceted comparative framework aiming to reveal the general rules in the observed patterns and underlying causes of elevational beta-diversity. We have found that: first, the turnover process dominated altitudinal patterns of species beta-diversity (ßsim > ßsne), whereas the nestedness process appeared relatively more important for elevational trait dissimilarity (ßfuncsim < ßfuncsne); second, the taxonomic turnover was relative higher than its phylogenetic and functional analogues (ßsim > ßphylosim/ßfuncsim), conversely, nestedness-resultant trait dissimilarity tended to be higher than the taxonomic and phylogenetic measures (ßfuncsne > ßsne/ßphylosne); and third, as elevational distance increased, the contradicting dynamics of environmental filtering and limiting similarity have jointly led the elevational patterns of beta-diversity, especially at taxonomic dimension. Based on these findings, we infer that the species turnover among phylogenetic relatives sharing similar functional attributes appears to be the main cause of shaping the altitudinal patterns of multi-dimensional beta-diversity. Owing to the methodological limitation in the randomization approach, currently, it remains extremely challenging to distinguish the influence of the neutral process from the offset between opposing niche-based processes. Despite the complexities and uncertainties during species assembling, with a multi-dimensional comparative perspective, this work offers us several important commonalities of elevational beta-diversity dynamics.

Effect of Ca(OH)2 on the Release Characteristics of HCl during Sludge Combustion.

With the addition of Ca(OH)2, the effects of combustion temperature, moisture, sludge particle size, and chlorine-containing additives on the removal of HCl during sludge combustion were studied. The experimental results showed that combustion temperature and moisture content promoted the formation of HCl and Ca(OH)2 played a key role in the formation of HCl during sludge combustion. Under the best conditions of a sludge particle size of 380-250 µm, moisture content of 5%, temperature of 850 °C, and Ca(OH)2/sludge weight ratio of 3/10, the HCl capture efficiency was 79.81%. In addition, the effect of PVC on the production of HCl was greater than that of NaCl, probably because the lattice energy of NaCl was much higher, indicating that inorganic chlorine was not the main source of HCl. Ca(OH)2 can effectively inhibit the formation of HCl, which had practical guiding significance for the formation of HCl during the sludge combustion, especially the sludge containing chlorine.

Study on the influencing factors of removal of NOX from cement kiln flue gas by sewage sludge as a denitration agent.

Experimental study on the influencing factors of using sewage sludge as a denitration agent for cement industry was carried out on a self-made laboratory-scale fluid-bed reactor. Results indicate that sludge combustion at 900 °C shows an ideal NOX (the sum of NO and NO2) removal activity under simulated working conditions of cement precalciner. The optimal removal efficiency of NOX can reach 70.36 ± 3.59% in the presence of cement raw meal (CRM) at a sludge particle size range of 0.18-0.25 mm and the sludge dosage of 0.75 g/min. Besides, the NOX removal efficiency increases to 76.94 ± 5.02% in the absence of CRM, indicating that cement raw meal inhibits the NOX removal. This phenomenon may be attributed to the fact that CRM has promotion effect on NH3 produced and obvious inhibitory effect on CO produced; while NH3 and CO play a leading role in NOX reduction, the combined effect leads to the decrease of NOX removal. Moreover, the relationship between the composition of CRM on the inhibition of NOX removal is MgO < CaCO3 < CRM < Al2O3

Development of an Enzyme-Mediated, Site-Specific Method to Conjugate Toll-Like Receptor 2 Agonists onto Protein Antigens: Toward a Broadly Protective, Four Component, Group A Streptococcal Self-Adjuvanting Lipoprotein-Fusion Combination Vaccine.

Subunit vaccines composed of protein antigens covalently attached to Toll-like receptor (TLR) agonists elicit superior immune responses compared to mixtures of antigens and TLR agonists. Among different conjugation approaches, enzyme-mediated ligation is one of the few that provides an opportunity for the generation of homogeneous, molecularly defined products in which protein antigens are maintained with native structures, which is most critical to elicit protective immune responses upon vaccination. Four highly conserved protein antigens from Group A Streptococcus (GAS) have the potential to be safe and efficacious vaccine candidates. After a TLR2 agonist fibroblast-stimulating lipopeptide-1 (FSL-1) was successfully attached onto each antigen using sortase A and techniques for their purification were developed, a combination vaccine containing interleukin 8 (IL-8) protease (Streptococcus pyogenes cell envelope proteinase [SpyCEP]), Group A Streptococcal C5a peptidase (SCPA), anchorless virulence factor arginine deiminase (ADI), and trigger factor (TF)-TLR2 conjugates was produced. This combination was assessed for immunity in mice and compared with mixtures of the four antigens with FSL-1 or alum. High titer antigen-specific IgG antibodies were detected from all vaccine groups, with antibodies elicited from FSL-1 conjugates around 10-fold higher compared to the FSL-1 mixture group. Furthermore, the FSL-1 conjugates afforded a more balanced TH1/TH2 immune response than the alum-adjuvanted group, suggesting that this combination vaccine represents a promising candidate for the prevention of GAS diseases. Thus, we established a conjugation platform that allows for the production of defined, site-specific antigen-adjuvant conjugates, which maintain the native three-dimensional structure of antigens and can be potentially applied to a variety of protein antigens.

Simultaneous Removal of SO2 and NO by O3 Oxidation Combined with Wet Absorption.

The effects of ozone concentration, NaOH concentration, type and concentration of additives, initial pH, temperature, and NO and SO2 concentration on simultaneous removal of NO and SO2 were studied through ozone oxidation combined with wet absorption. Results indicated that ozone concentration and the type and concentration of additives had the most significant effect on NO removal. The optimal ozone concentration was 250 ppm (NO/NO2 = 1), and the best additive was KMnO4. The removal efficiency of NO x was as high as 97.86% when NO/NO2 = 1, and the concentration of KMnO4 was 0.025 mol/L. Considering economic and other factors, the KMnO4 concentration was selected to be 0.006 mol/L. At this time, the removal efficiencies of NO x and SO2 were 81.35 and 100%, respectively. This method has potential application prospects for simultaneous removal of SO2 and NO in the industrial flue gas.

A Self-Adjuvanting Vaccine Platform: Optimization of Site-Specific Sortase A Mediated Conjugation of Toll-Like Receptor 2 Ligands onto the Carboxyl or Amino terminus of Recombinant Protein Antigens.

Self-adjuvanting vaccines, consisting of recombinant protein antigens and covalently attached Toll-like receptor (TLR) agonists, have the ability to simultaneously and efficiently deliver antigen and TLR adjuvant to antigen presenting cells (APCs). Here, an enzyme-mediated ligation approach was used to overcome difficulties in producing homogeneous, molecularly defined self-adjuvanting vaccine products under native conditions. This process was optimized to allow the incorporation of the lipopeptide TLR2 agonist fibroblast-stimulating lipopeptide (FSL)-1 onto the N- or C-termini of recombinant protein antigens, employing the enzyme Staphylococcus aureus sortase A (SrtAsa) penta mutant. In addition, because SrtAsa-mediated ligations are reversible, a tryptophan zipper derived sequence was introduced into both reactants, which was demonstrated to improve ligation efficiency through the formation of a ß-hairpin structure that hinders the reverse reaction. Finally, it was demonstrated that N- or C-terminal conjugation, and the incorporation of the ß-hairpin structure, did not affect the TLR2-agonist activities of protein antigen TLR agonist conjugates. Overall, this SrtAsa-mediated ligation platform enabled production of antigen TLR2 agonist conjugates with enhanced ligation efficiency, with the conjugates demonstrating potent TLR2 signaling activation (EC50 <1nM).

Semisynthetic, self-adjuvanting vaccine development: Efficient, site-specific sortase A-mediated conjugation of Toll-like receptor 2 ligand FSL-1 to recombinant protein antigens under native conditions and application to a model group A streptococcal vaccine.

Protein antigens are, in general, weakly immunogenic, and therefore require co-delivery with adjuvants to stimulate potent immune responses. The fusion of (poly)peptide antigens to immunostimulatory adjuvants (e.g. Toll-like receptor (TLR) agonists) has been demonstrated to greatly improve vaccine potency compared to mixtures of antigen and adjuvant. Chemical approaches, to enable the rapid, site-specific and high-yielding linkage of TLR2 ligands to recombinant protein antigens, have been previously optimized. These approaches require the use of denaturing conditions to ensure high reaction yields, which limits their application, as maintenance of native protein folding is necessary to elicit antibodies against conformational epitopes. Here, this work aimed to optimize an alternative method, to ensure the efficient bioconjugation of TLR2 ligands onto folded protein antigens. An enzyme-mediated approach, using Staphylococcus aureus sortase A (or a penta mutant with enhanced efficiency), was optimized for reaction yield and time, as well as enzyme type and amount. This approach enabled the site-specific conjugation of the TLR2-agonist fibroblast-stimulating lipopeptide-1 (FSL-1) onto a model group A Streptococcus (GAS) recombinant polytope antigen under conditions that maintain protein folding, yielding a homogeneous, molecularly-defined product, with ligation yields as high as 90%. Following intramuscular (IM) administration of the ligation product to humanized plasminogen AlbPLG1 mice, high-titer, antigen-specific IgG antibodies were observed, which conferred protection against subcutaneous challenge with GAS strain 5448. In comparison, mixtures of the GAS antigen with aluminum hydroxide or FSL-1 failed to provide protection, with the FSL-1 mixture yielding ~1000-fold lower antigen-specific IgG antibody titers, and the mixture with alum yielding a Th2-biased response compared to the more balanced Th1/Th2 responses observed with the FSL-1 conjugate. Overall, a FSL-1 bioconjugation method for the efficient production of antigen-TLR2 agonist conjugates, which maintain protein folding, was produced, with broad utility for the development of self-adjuvanting vaccines against subunit protein antigens.

Night-light Image Restoration Method Based on Night Scattering Model for Luojia 1-01 Satellite.

Aiming at solving the degradation problem of Luojia 1-01 night-light remote sensing images, the main reason for the "glow" phenomenon was analyzed. The APSF (Atmospheric Point Spread Function) template of night-light image was obtained from atmospheric source scattering. The template was used as the initial value in the regularization restoration model in this paper. Experiments were carried out using single point and regional images. The results demonstrate that the estimated APSF and restoration results of the method are better than those from other methods, and the image quality is improved after restoration.

Taxonomic and Functional Ant Diversity Along tropical, Subtropical, and Subalpine Elevational Transects in Southwest China.

Although elevational gradients of biodiversity have long been the topic of scientific research, information on patterns of, and processes that shape insect community structure across elevation is still lacking. Addressing this gap requires the use of both taxonomic and functional approaches when studying diversity across elevational gradients. In this study, we examined taxonomic and functional alpha and beta diversity of ant assemblages sampled along tropical, subtropical, and subalpine elevational transects in Yunnan Province, southwest China. Species richness was used to quantify taxonomic alpha diversity, and two indices (FD and FRic) were calculated using morphological measurements to quantify functional alpha diversity. Taxonomic and functional beta diversity were partitioned into their turnover- and nestedness-resultant components. Though temperature and functional alpha diversity decreased linearly with increasing elevation, taxonomic alpha diversity showed a significant logarithmic decrease, with few species present at elevations greater than 3000 m a.s.l. The turnover-resultant component of taxonomic beta diversity increased with increasing elevational distance, while the nestedness-resultant component of functional beta diversity increased with increasing elevational distance in the subtropical transect. The observed patterns of taxonomic and functional diversity reflected ants' thermophilic nature, implying functional adaptations (i.e., nested functional diversity) at higher elevations where environmental conditions were unfavorable.

Systematics of the ant genus Proceratium Roger (Hymenoptera, Formicidae, Proceratiinae) in China - with descriptions of three new species based on micro-CT enhanced next-generation-morphology.

The genus Proceratium Roger, 1863 contains cryptic, subterranean ants that are seldom sampled and rare in natural history collections. Furthermore, most Proceratium specimens are extremely hairy and, due to their enlarged and curved gaster, often mounted suboptimally. As a consequence, the poorly observable physical characteristics of the material and its scarcity result in a rather challenging alpha taxonomy of this group. In this study, the taxonomy of the Chinese Proceratium fauna is reviewed and updated by combining examinations of traditional light microscopy with x-ray microtomography (micro-CT). Based on micro-CT scans of seven out of eight species, virtual 3D surface models were generated that permit in-depth comparative analyses of specimen morphology in order to overcome the difficulties to examine physical material of Proceratium. Eight Chinese species are recognized, of which three are newly described: Proceratium bruelheidei Staab, Xu & Hita Garcia, sp. n. and P. kepingmaisp. n. belong to the P. itoi clade and have been collected in the subtropical forests of southeast China, whereas P. shoheisp. n. belongs to the P. stictum clade and it is only known from a tropical forest of Yunnan Province. Proceratium nujiangense Xu, 2006 syn. n. is proposed as a junior synonym of P. zhaoi Xu, 2000. These taxonomic acts raise the number of known Chinese Proceratium species to eight. In order to integrate the new species into the existing taxonomic system and to facilitate identifications, an illustrated key to the worker caste of all Chinese species is provided, supplemented by species accounts with high-resolution montage images and still images of volume renderings of 3D models based on micro-CT. Moreover, cybertype datasets are provided for the new species, as well as digital datasets for the remaining species that include the raw micro-CT scan data, 3D surface models, 3D rotation videos, and all light photography and micro-CT still images. These datasets are available online (Dryad, Staab et al. 2018, http://dx.doi.org/10.5061/dryad.h6j0g4p).

Bioconjugation Approaches to Producing Subunit Vaccines Composed of Protein or Peptide Antigens and Covalently Attached Toll-Like Receptor Ligands.

Traditional vaccines derived from attenuated or inactivated pathogens are effective at inducing antibody-based protective immune responses but tend to be highly reactogenic, causing notable adverse effects. Vaccines with superior safety profiles can be produced by subunit approaches, utilizing molecularly defined antigens (e.g., proteins and polysaccharides). These antigens, however, often elicit poor immunological responses, necessitating the use of adjuvants. Immunostimulatory adjuvants have the capacity to activate antigen presenting cells directly through specific receptors (e.g., Toll-like receptors (TLRs)), resulting in enhanced presentation of antigens as well as the secretion of proinflammatory chemokines and cytokines. Consequently, innate immune responses are amplified and adaptive immunity is generated. Recently, site-specific conjugation of such immunostimulatory adjuvants (e.g., TLR ligands) onto defined antigens has shown superior efficacy over unconjugated mixtures, suggesting that the development of chemically characterized immunostimulatory adjuvants and optimized approaches for their conjugation with antigens may provide a better opportunity for the development of potent, novel vaccines. This review briefly summarizes various TLR agonists utilized as immunostimulatory adjuvants and focuses on the development of techniques (e.g., recombinant, synthetic, and semisynthetic) for generating adjuvant-antigen fusion vaccines incorporating peptide or protein antigens.

An outbreak of norovirus gastroenteritis associated with a secondary water supply system in a factory in south China.

BACKGROUND: Between September 17 and October 3, 2009, hundreds of workers employed in a manufacturing factory in Shenzhen, a city in south China developed a sudden onset of acute gastroenteritis. A retrospective cohort study is designed to identify the risk factors and control this outbreak. METHODS: Information on demographic characteristics, working place, the history of contact with a person having diarrhea and/or vomiting, drink water preference and frequency, eating in the company cafeteria or outside the company, hand-washing habits and eating habits is included. Furthermore, in order to find the contamination source, we investigated the environment around the underground reservoir and collected water samples from the junction between municipal supply water system and underground reservoir to test potential bacteria and virus, examine the seepage tracks on the wall of the underground reservoir from the side of septic tank, and check the integrity and attitude of this lid. Relative risk was presented and Chi-square test was performed. All the analyses were performed with OpenEpi software version 2.3.1 online. RESULTS: The cohort study demonstrated that the workers who had direct drink water were 3.0 fold more likely to suffer from acute gastroenteritis than those who consumed commercial bottled water. The direct drinking water, water of the tank of buildings, and the underground reservoir were positive only for norovirus. Norovirus was also detected from stool and rectal swab samples from patients with acute gastroenteritis. The underground reservoir was found to be the primary contamination source. Further environmental investigation showed that the norovirus contaminated substance entered into the underground reservoir via access holes in lid covering this underground reservoir. CONCLUSION: This acute gastroenteritis outbreak was caused by the secondary supply system contaminated by norovirus in this factory. The outbreak of gastroenteritis cases caused by norovirus frequently occurred in China due to a lack of surveillance and supervision, and due to faults in the construction of such water systems. Therefore, more attentions should pay to the secondary supply water system in China.

Rapid and simultaneous detection of Ureaplasma parvum and Chlamydia trachomatis antibodies based on visual protein microarray using gold nanoparticles and silver enhancement.

Based on gold-labeled silver stain (GLSS) method, we developed the visual protein microarray for simultaneous, sensitive, and specific detection of Ureaplasma parvum and Chlamydia trachomatis using N-terminus multiple-banded antigen (NMBA) of U. parvum and major outer membrane protein of C. trachomatis. The specific antigens were immobilized on glass surface that was treated with 3-glycidoxypropyltrimethoxysilane, and they were used as the capturing probes to recognize the complementary target antibodies binding to the detecting probes of Nano-gold-Staphylococcal protein A (SPA). In the "sandwich" format, Nano-gold-SPA probe was used as an indicator and GLSS was applied to amplify the detection signals and produce black image on array spots, which were visible with naked eyes. In our model arrays, the detection limit of protein microarray was as low as 2 ng/mL, and the lowest titer of detectable antibody was 1:128; thus, this sensitivity was comparable to the fluorescent detection method. The visual simultaneous protein microarrays were used to detect total 186 clinical samples, which had been determined by enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative real-time polymerase chain reaction; the results were identical and no distinct difference (P > 0.05) existed between them. Our results demonstrate that we have developed the visual protein microarray technique, which is of high sensitivity and high specificity, and it may have potential in clinical applications.