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Background: Plants that contain brewing microorganisms are used in traditional fermentation starters, which are an essential part of local diet, nutrition, life, and health. Regionally, the plant species used and the microorganisms included in traditional fermentation starters are diverse, endowing local fermented drinks with different flavors and health benefits. However, related traditional knowledge has been scarcely documented or revealed. Methods: An ethnobotanical survey was conducted in five towns of Nayong County in northwestern Guizhou, China. Snowball sampling, semi-structured interviews, free lists, and participatory observation were used to collect information on Jiuqu Plants (JPs) and jiuqu-making techniques. The PacBio platform was used to study the microbial community structure and diversity in the Chuanqing people's jiuqu. Results: In total, 225 informants were interviewed, including 116 who provided plants and technological processes for making Chinese baijiu jiuqu (CBJ) and 139 who provided information about making fermented glutinous rice jiuqu (FGRJ). This study found that older people have more abundant knowledge about CBJ plants. Poaceae was found to be the dominant family used in making CBJ and FGRJ (7 species each). Compared to individual plant parts, the whole plant is most commonly used in two kinds of jiuqu (19.5% in CBJ and 22.6% in FGRJ). The Chuanqing people's jiuqu is used to treat dietary stagnation and indigestion. The highest relative frequency of citation of the CBJ plant was Ficus tikoua Bureau, and the counterpart of the FGRJ plant was Buddleja macrostachya Benth. The dominant bacterial species in jiuqu were Gluconobacter japonicus (YQ1, YQ4) and Pediococcus pentosaceus (YQ2, YQ3), and the dominant fungal species was Rhizopus oryzae. Conclusion: For the first time, this study documents the unique traditional jiuqu knowledge and reveals the microbial mystery behind the FGRJ of the Chuanqing people. Therefore, this study encourages the use of online social media platforms in order to spread Jiuqu culture, the use of the new media wave in order to create multimedia databases, and also suggests that local communities should develop preservation intervention programs, in addition to nurturing the inheritors in order to prevent the disappearance of traditional Jiuqu knowledge. This research contributes to the conservation and demystification of the traditional jiuqu knowledge of the Chuanqing people and lays the foundation for further research on its microbiology, nutrition, and metabolomics. [ABSTRACT FROM AUTHOR]

Background Hemoglobin (Hb) Hekinan II (A27; Glu-Asp) is an α-chain variant, and its interaction with the common Southeast Asian (--SEA/) α-thalassemia (α-thal) deletion is rarely reported. This study provides a clinical update of Hb Hekinan II associated with (--SEA/) α-thal. Methods A total of 11 simple heterozygotes and 20 composite heterozygotes for Hb Hekinan II and (--SEA/) α-thal were included based on molecular diagnosis. Results Hb Hekinan II exhibited a significant increase in hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin content, but a decrease in red blood cell level compared with α+ thalassemia deletion. Compared with (--SEA/) α-thal, composite heterozygotes for Hb Hekinan II and (--SEA/) α-thal showed similar erythrocyte parameters. Both heterozygotes with and without (--SEA/) α-thal showed low Hb A2 level. Hb Hekinan II showed abnormal performance in high-performance liquid chromatography but not in capillary electrophoresis. Conclusion Hb Hekinan II is a benign Hb variant. The heterozygotes exhibit clinically asymptomatic coinheritance with (--SEA/) α-thal having comparable hematological phenotype to simple (--SEA/) α-thal. The combination of hematological and molecular analysis helped to improve the detection rate of this rare variant. [ABSTRACT FROM AUTHOR]

Stroke has become a major cause of death and disability worldwide. The cellular recycling pathway autophagy has been implicated in ischemia-induced neuronal changes, but whether autophagy plays a beneficial or detrimental role is controversial. Hydroxysafflor Yellow A (HSYA), a popular herbal medicine, is an extract of Carthamus tinctorius and is used to treat ischemic stroke (IS) in China. HSYA has been shown to prevent cardiovascular and cerebral ischemia/reperfusion injury in animal models. However, the specific active ingredients and molecular mechanisms of HSYA in IS remain unclear. Here, we investigated the effect of HSYA treatment on autophagy in a rat model of IS. IS was induced in rats by middle cerebral artery occlusion. Rats were treated once daily for 3 days with saline, HYSA, or the neuroprotective agent Edaravone. Neurobehavioral testing was performed on days 1, 2, and 3 post-surgery. Brains were removed on day 3 post-surgery for histological evaluation of infarct area, morphology, and for qRT-PCR and western blot analysis of the expression of the autophagy factor LC3 and the signaling molecules HIF-1 α , BNIP3, and Notch1. Molecular docking studies were performed in silico to predict potential interactions between HSYA and LC3, HIF-1 α , BNIP3, and Notch1 proteins. The result showed that HSYA treatment markedly alleviated IS-induced neurobehavioral deficits and reduced brain infarct area and tissue damage. HSYA also significantly reduced hippocampal expression levels of LC3, HIF-1 α , BNIP3, and Notch1. The beneficial effect of HSYA was generally superior to that of Edaravone. Molecular modeling suggested that HSYA may bind strongly to HIF-1 α , BNIP3, and Notch1 but weakly to LC3. In conclusion, HSYA inhibits post-IS autophagy induction in the brain, possibly by suppressing HIF-1 α , BNIP3 and Notch1. HSYA may have utility as a post-IS neuroprotective agent. [ABSTRACT FROM AUTHOR]

Poultries including chickens, ducks, geese, and pigeons are widely used in the biological and medical research in many aspects. The genetic quality of experimental poultries directly affects the results of the research. In this study, following electrophoresis analysis and short tandem repeat (STR) scanning, we screened out the microsatellite loci for determining the genetic characteristics of Chinese experimental chickens, ducks, geese, and pigeons. The panels of loci selected in our research provide a good choice for genetic monitoring of the population genetic diversity of Chinese native experimental chickens, ducks, geese, and ducks. [ABSTRACT FROM AUTHOR]

Background: The current research was to assess the relationship of the solute carrier family 44 member 4 (SLC44A4) rs577272, notch receptor 4 (NOTCH4) rs3134931 SNPs and serum lipid levels in the Han and Maonan ethnic groups. Methods: The genetic makeup of the SLC44A4 rs577272 and NOTCH4 rs3134931 SNPs in 2467 unrelated subjects (Han, 1254; Maonan,1213) was obtained by using polymerase chain reaction and restriction fragment length polymorphism technique, combined with gel electrophoresis, and confirmed by direct sequencing. Results: The genotype frequencies of SLC44A4 rs577272 and NOTCH4 rs3134931 SNPs were different between Han and Maonan populations (P < 0.05); respectively. The SLC44A4 rs577272 SNP was associated with total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) levels in Maonan group. The NOTCH4 rs3134931 SNP was associated with triglyceride (TG) in Han; and TG and low-density lipoprotein cholesterol (LDL-C) levels in Maonan groups (P < 0.025–0.001). Stratified analysis according to gender showed that the SLC44A4 rs577272 SNP was associated with TC and HDL-C in Han and Maonan females; TC in Maonan males, meanwhile, the NOTCH4 rs3134931 SNP was associated with TG and HDL-C in Han males; TG in Han females; TG and LDL-C in Maonan males; and TG, HDL-C and LDL-C in Maonan females. Linkage disequilibrium analysis showed that the most common haplotype was rs577272G-rs3134931A (> 50%) in both Han and Maonan groups. The haplotype of rs577272G-rs3134931A was associated with TG and HDL-C in Han; and TC, TG and HDL-C in Maonan ethnic groups. Conclusions: These results suggest that the relationship among SLC44A4 rs577272, NOTCH4 rs3134931 SNPs and serum lipid parameters may vary depending on the gender and/or ethnicity/race in some populations. Haplotypes could explain more changes in serum lipid parameters than any single SNP alone particularly for TC, TG and HDL-C. [ABSTRACT FROM AUTHOR]

Objectives: To characterize the oxazolidinone resistance gene poxtA on broad-host-range Inc18 plasmids from CC17 Enterococcus faecium of pig origin.Methods: Oxazolidinone-resistant E. faecium isolates were screened for the presence of poxtA. The poxtA-carrying isolates were characterized by antimicrobial susceptibility testing, conjugation, S1-PFGE and hybridization. The poxtA-carrying plasmids were completely sequenced and their instability was verified.Results: Two individual CC17 E. faecium strains were positive for poxtA. S1-PFGE and hybridization revealed the presence of a poxtA-carrying plasmid of ∼62 kb in both WZ27-2 and the transconjugant, while poxtA-carrying plasmids of different sizes were observed in QF25-1 and the transconjugant. The two poxtA-carrying plasmids, pC25-1 and pC27-2, belonged to the broad-host-range plasmids of the Inc18 family and carried dfrG, aadE, Δsat4, aph(3')-III, erm(B), tet(M), tet(L) and fexB. Plasmid pC27-2 was virtually identical to pC25-1, with minor differences. The calculated transfer frequency was ∼0.87 × 10-8 and ∼1.03 × 10-7 per recipient to plasmids pC25-1 and pC27-2, respectively. Instability assays of the region with four adjacent IS1216Es, which forms three IS1216E translocatable units, revealed the formation of a series of mosaic circular intermediates.Conclusions: We report the emergence of the plasmid-mediated oxazolidinone resistance gene poxtA in E. faecium from different farms in China. Comparison of the poxtA genetic context suggests that IS1216E elements play an important role in the dissemination of poxtA. The co-occurrence of poxtA with other antimicrobial and heavy metal resistance genes on the broad-host-range plasmids of the Inc18 family may lead to the co-selection of poxtA, contributing to its persistence and accelerating its dissemination. [ABSTRACT FROM AUTHOR]

Objectives: To investigate the occurrence, the genetic environment and the functionality of novel variants of the MDR gene cfr(C) in Campylobacter from China.Methods: A total of 370 Campylobacter isolates of porcine and chicken origin collected from three regions of China in 2015 were screened for cfr(C) by PCR. The phenotypes and genotypes of cfr(C)-positive isolates were investigated by antimicrobial susceptibility testing, PFGE, MLST, S1-PFGE, Southern blotting and WGS. Quantitative RT-PCR was used to compare the expression levels of the cfr(C) variants in their original isolate and clone constructs in Campylobacter jejuni NCTC 11168.Results: Four (1.1%) porcine Campylobacter coli isolates were positive for cfr(C). They failed to show elevated MICs of phenicols. The deduced Cfr(C) sequences identified exhibited 2-6 amino acid changes compared with the original Cfr(C) reported in the USA. Cloning of the cfr(C) variant genes into C. jejuni NCTC 11168 resulted in ≥32-fold increases in the MICs of phenicols, indicating that the cfr(C) variant genes are functional. The cfr(C)-carrying isolates belonged to three genotypes and WGS analysis revealed the cfr(C) genes were chromosomally located in MDR genomic islands, which contained multiple antibiotic resistance genes of Gram-positive origin.Conclusions: This study identified chromosomal cfr(C) genes in C. coli isolates from China. They appeared functionally dormant in the original isolates but were fully functional when cloned and expressed in C. jejuni. The cfr(C) genes were co-transferred with other antibiotic resistance genes, possibly from Gram-positive bacteria. These findings reveal new insights into the function and transmission of cfr(C) in Campylobacter. [ABSTRACT FROM AUTHOR]

Introduction: Fanconi anemia (FA), as one of the congenital bone marrow failure syndromes, is characterized by severe bone marrow hypocellularity and pancytopenia which is similar with acquired aplastic anemia (AAA). However, patients with FA or AAA need an accurate diagnose, as the two syndromes differ significantly in both treatment and prognosis. FA results from gene mutations of the FA pathway genes specifically required for DNA repair, and the mutation of these genes contributes to the genome instability of FA cells. Based on this feature, chromosome aberration (CA) has been used as a "golden standard" to the auxiliary diagnosis of FA from AAA. However, CA diagnose calls for more technical requirements and a long time for the subsequent statistical analysis. Methods: In our study, another two genome instability examination tools, cytokinesis‐block micronucleus (CBMN) and single‐cell gel electrophoresis (SCGE), were used to distinguish FA patients from AAA patients, compared with CA. Results: The results suggested that significant differences were observed in the FA patients compared with the AAA patients and the controls using all of the three genomic instability examination tools. However, CBMN is the most cost‐effective method to distinguish FA patients from AAA patients among the three genome instability examination tools, when the time costs, instrument costs, and technical costs were compared. Conclusion: In areas with economic and technical limitations, CBMN is an alternative assay to help distinguish FA patients from AAA patients. [ABSTRACT FROM AUTHOR]

Objectives: To investigate the presence and molecular characteristics of oxazolidinone resistance genes cfr and optrA in staphylococci from household animals in rural China.Methods: Various samples were collected from household animals in 12 rural villages. Staphylococcal isolates showing florfenicol MICs ≥10 mg/L were identified and screened for the presence of cfr and/or optrA. PCR-positive isolates were characterized by antimicrobial susceptibility testing, S1 nuclease PFGE and Southern blotting. WGS data were analysed to identify the core-genome phylogenetic profile of each isolate as well as the genetic environment of cfr and/or optrA.Results: Nine optrA-positive (seven Staphylococcus sciuri and two Staphylococcus simulans) and 10 cfr-positive staphylococci were identified from eight and five villages, respectively. The gene optrA was chromosomally encoded in all nine isolates, whereas cfr was located on a plasmid in one S. sciuri and three Staphylococcus saprophyticus and in the chromosomal DNA of single Staphylococcus cohnii and Staphylococcus lentus isolates and two S. sciuri isolates. The remaining two cfr-carrying Staphylococcus haemolyticus isolates were indistinguishable by PFGE. Most optrA- or cfr-carrying staphylococci also harboured phenicol, tetracycline and/or macrolide-lincosamide-streptogramin B resistance genes. Genetic environment analysis showed that, for the first time, optrA was associated with transposon Tn6261, while cfr was adjacent to both a tnp (transposase) gene and a Tn558 transposon.Conclusions: The current study reveals for the first time the wide distribution of oxazolidinone resistance genes optrA and cfr in household animals in rural areas of China and is the first identification of optrA in S. simulans isolates. [ABSTRACT FROM AUTHOR]

Tooth agenesis is one of the most common developmental anomalies affecting function and esthetics. The paired-domain transcription factor, Pax9, is critical for patterning and morphogenesis of tooth and taste buds. Mutations of PAX9 have been identified in patients with tooth agenesis. Despite significant progress in the genetics of tooth agenesis, many gaps in knowledge exist in refining the genotype-phenotype correlation between PAX9 and tooth agenesis. In the present study, we complete genetic and phenotypic characterization of multiplex Chinese families with nonsyndromic (NS) tooth agenesis. Direct sequencing of polymerase chain reaction products revealed 9 novel (c.140G>C, c.167T>A, c.332G>C, c.194C>A, c.271A>T, c.146delC, c.185_189dup, c.256_262dup, and c.592delG) and 2 known heterozygous mutations in the PAX9 gene among 120 probands. Subsequently, pedigrees were extended, and we confirmed that the mutations co-segregated with the tooth agenesis phenotype (with exception of families in which DNA analysis was not available). In 1 family ( n = 6), 2 individuals harbored both the PAX9 c.592delG mutation and a heterozygous missense mutation (c.739C>T) in the MSX1 gene. Clinical characterization of families segregating a PAX9 mutation reveal that all affected individuals were missing the mandibular second molar and their maxillary central incisors are most susceptible to microdontia. A significant reduction of bitter taste perception was documented in individuals harboring PAX9 mutations ( n = 3). Functional studies revealed that PAX9 haploinsufficiency or a loss of function of the PAX9 protein underlies tooth agenesis. [ABSTRACT FROM AUTHOR]

Objectives: This study identified and characterized mcr-1-positive Enterobacteriaceae (MCRPE) and carbapenemase-producing Enterobacteriaceae (CPE) in hospital sewage water.Methods: Influent and effluent sewage samples were collected from five tertiary hospitals in Beijing in December 2016. Samples were screened for MCRPE and CPE using antibiotic selection media. Results were confirmed by PCR amplification of β-lactamase and colistin resistance (mcr-1 and mcr-2) genes and by sequencing. Antimicrobial susceptibility testing, MLST, conjugation and plasmid typing and S1-nuclease-PFGE/Southern blotting were performed for all MCRPE and CPE isolates.Results: Nine MCRPE and 12 CPE isolates were obtained. All mcr-1-positive isolates (n = 9) were Escherichia coli and belonged to eight different STs. The blaKPC-2-positive Enterobacteriaceae included Klebsiella pneumoniae (n = 4), Enterobacter cloacae (n = 4) and Citrobacter freundii (n = 1) isolates. Two C. freundii isolates and one E. cloacae isolate harboured the blaNDM-1 gene. MLST analysis revealed distinct genetic relatedness among all ST11 K. pneumoniae but not among any other carbapenemase-producing isolates. Conjugation and plasmid typing confirmed that three MCRPE isolates harboured mcr-1 on the self-transmissible IncX4 plasmid and the blaNDM-1 gene on the IncX3 plasmid. The sizes of the plasmids harbouring mcr-1, blaNDM-1 and blaKPC-2 were ∼33 to ∼240, ∼40 to ∼75 and ∼30 to ∼90 kb, respectively.Conclusions: To the best of our knowledge, this is the first report of mcr-1-positive E. coli and blaNDM-1-carrying E. cloacae and C. freundii in hospital sewage water. These findings, especially the diversity of MCRPE and K. pneumoniae ST11 that harbour the blaKPC-2 gene, suggest that monitoring and management of hospital sewage water should be enhanced. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the contribution of SHISA3 promoter methylation to laryngeal squamous cell carcinoma (LSCC). SHISA3 promoter methylation status and expression were determined using methylation-specific polymerase chain reaction (MSP) and quantitative real-time PCR (qRT-PCR) in 93 paired LSCC and adjacent normal tissues, respectively. Furthermore, the regulatory function of the SHISA3 promoter fragment was analyzed using a luciferase reporter assay. The results reveal that there is a significant increase in SHISA3 methylation in LSCC tissues compared with corresponding nontumor tissues P=4.58E-12. The qRT-PCR results show a significant association between SHISA3 methylation and expression in LSCC P=1.67E-03. In addition, the area under the receiver operating characteristic curve was 0.91. Consequently, a log-rank test and multivariate Cox analysis suggest that SHISA3 promoter hypermethylation is a predictor of poor overall survival for LSCC (log-rank P = 0.024; HR = 2.71; 95% CI = 1.024–7.177; P = 0.047). The results indicate that SHISA3 promoter hypermethylation might increase the risk of LSCC through regulation of gene expression and is a potential diagnostic and prognostic biomarker for LSCC. [ABSTRACT FROM AUTHOR]

The 41 kD flagellin of Borrelia burgdorferi (B. burgdorferi) is a major component of periplasmic flagellar filament core and a good candidate for serodiagnosis in early stage of Lyme disease. Here, we chose 89 B. burgdorferi strains in China, amplified the gene encoding the 41 kD flagellin, and compared the sequences. The results showed that genetic diversity presented in the 41 kD flagellin genes of all 89 strains among the four genotypes of B. burgdorferi, especially in the genotype of B. garinii. Some specific mutation sites for each genotype of the 41 kD flagellin genes were found, which could be used for genotyping B. burgdorferi strains in China. Human B-cell epitope analysis showed that thirteen of 15 nonsynonymous mutations occurred in the epitope region of 41 kD flagellin and thirty of 42 B-cell epitopes were altered due to all 13 nonsynonymous mutations in the epitope region, which may affect the function of the antigen. Nonsynonymous mutations and changed human B-cell epitopes exist in 41 kD flagellin of B. burgdorferi sensu lato strains; these changes should be considered in serodiagnosis of Lyme disease. [ABSTRACT FROM AUTHOR]

The metal pollution has posed great risk on the coastal organisms along the Jiulongjiang Estuary in South China. In this work, two-dimensional electrophoresis-based proteomics was applied to the oysters Crassostrea hongkongensis from metal pollution sites to characterize the proteomic responses to metal pollution. Metal accumulation and proteomic responses indicated that the oysters from BJ site were more severely contaminated than those from FG site. Compared with those oyster samples from the clean site (JZ), metal pollution induced cellular injuries, oxidative and immune stresses in oyster heapatopancreas from both BJ and FG sites via differential metabolic pathways. In addition, metal pollution in BJ site induced disturbance in energy and lipid metabolisms in oysters. Results indicated that cathepsin L and ferritin GF1 might be the biomarkers of As and Fe in oyster C. hongkongensis , respectively. This study demonstrates that proteomics is a useful tool for investigating biological effects induced by metal pollution. [ABSTRACT FROM AUTHOR]

Pulmonary diseases caused by nontuberculous mycobacteria (NTM) are increasing in incidence and prevalence worldwide. In this study, we identified NTM species of the clinical isolates from 8 provinces in China, in order to preliminarily provide some basic scientific data in the different species and distribution of NTM related to pulmonary disease in China. A total of 523 clinical isolates from patients with tuberculosis (TB) diagnosed clinically from 2005 to 2012 were identified to the species using conventional and molecular methods, including multilocus PCR, rpoB and hsp65 PCR-PRA, hsp65, rpoB, and 16S-23S internal transcribed spacer region sequencing. The isolates were identified into 3 bacterium genera, including NTM, Gordonia bronchialis, and Nocardia farcinica, and, for the 488 NTM isolates, 27 species were identified. For all the 27 species of NTM which were found to cause pulmonary infections in humans, the most prevalent species was M. intracellulare, followed by M. avium and M. abscessus. And seven other species were for the first time identified in patients with TB in China. NTM species identification is very important for distinguishing between tuberculosis and NTM pulmonary diseases, and the species diversity drives the creation of diverse and integrated identification methods with higher accuracy and efficacy. [ABSTRACT FROM AUTHOR]

Background: Malocclusion is a common disease of oral and maxillofacial region. The study was aimed to investigate levels changes of periodontal pathogens in malocclusion patients before, during and after orthodontic treatments, and to confirm the difference between adults and children. Method: One hundred and eight malocclusion patients (46 adults and 62 children at the school-age) were randomly selected and received orthodontic treatment with fixed orthodontic appliances. Subgingival plaques were Porphyromonas gingivalis (P.gingivalis), Fusobacterium nucleatum (F. nucleatum), Prevotella intermedia (P. intermedia) and Tannerella forsythensis (T. forsythensis) collected from the observed regions before and after treatment. Clinical indexes, including plaque index (PLI), gingival index (GI), sulcus bleeding index (SBI), probing depth (PD) and attachment loss (AL) of observed teeth were examined. Results: The detection rates of P.gingivalis, F. nucleatum, P. intermedia and T. forsythensis increased from baseline to the third month without significant difference, and then returned to pretreatment levels 12 month after applying fixed orthodontic appliances. Adults' percentage contents of P.gingivalis, F. nucleatum, P. intermedia and T. forsythensis were significantly higher than those of children at baseline and the first month, but not obvious at the third month. PLI and SBI were increased from baseline to the first and to the third month both in adults and children groups. Besides, PD were increased from baseline to first month, followed by a downward trend in the third month; however, all patients were failed to detect with AL. Conclusions: Periodontal and microbiological statuses of malocclusion patients may be influenced by fixed orthodontic appliances in both adults and children, more significant in children than in adults. Some microbiological indexes have synchronous trend with the clinical indexes. Long-term efficacy of fixed orthodontic appliances for malocclusion should be confirmed by future researches. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is an obligate intracellular parasitic protozoan that can infect almost all species of warm-blooded animals. As any chemical-based drugs could not act against the tissue cyst stage of T. gondii, vaccination may be one of the ideal control strategies. In the present study, two new vaccine candidates, named TgENO2 and TgTrxLp, were purified from Escherichia coli with pET-30a(+) expression system and then were injected into BALB/c mice to evaluate the protective efficacy against acute and chronic toxoplasmosis. The results showed that both the recombinant proteins, either alone or in combination, could elicit strong humoral and cellular immune responses with a higher level of IgG antibodies, IFN-γ, IL-2, CD4+, and CD8+ T cells as compared to those in mice from control groups. After acute challenge with tachyzoites of the GJS strain, mice immunized with rTgTrxLp (8±2.77 d), rTgENO2 (7.4±1.81 d), and rTgTrxLp + rTgENO2 (8.38±4.57 d) proteins showed significantly longer survival time than those that received Freund’s adjuvant (6.78±2.08 d) and PBS (6.38±4.65 d) (χ2 = 9.687, df = 4, P=0.046). The protective immunity of rTgTrxLp, rTgENO2, and rTgTrxLp + rTgENO2 proteins against chronic T. gondii infection showed 69.77%, 58.14%, and 20.93% brain cyst reduction as compared to mice that received PBS. The present study suggested that both TgENO2 and TgTrxLp were potential candidates for the development of multicomponent vaccines against toxoplasmosis. [ABSTRACT FROM AUTHOR]

A water-soluble polysaccharide (named RAP) was newly isolated from the stems ofRubus amabilis. Structural confirmation of the polysaccharide was provided by hydrolysis, periodate oxidation, Smith degradation, and methylation analysis, combined with nuclear magnetic resonance (NMR), capillary electrophoresis (CE), infrared spectroscopy (IR), and gas chromatography-mass spectra (GC-MS).In vitroimmunological enhancement activity was characterized using the proliferative activity of spleen lymphocytes and phagocytic activity of peritoneal macrophages in mice. The polysaccharide was mainly composed of xylose, arabinose, glucose, rhamnose, galactose, mannose, glucuronic acid, and galactocuronic acid in the molar ratio of 1.0:6.9:0.8:1.1:6.9:0.3:0.5:3.3, with the average molecular weight of 26.2 kDa. The linkage types of netural monosaccharides were as follows: the arabinose was →2) Ara (1→ and galactose were Gal (1→, →3) Gal (1→, →3,6) Gal (1→, →2,3,6) Gal (1→ and →2,3,6) Galf(1→. Xyl (1→, →6) Glc (1→, →2) Glc (1→, →3) Rha (1→, Rha (1→ and Man (1→ were also found in the structure. RAP-B-2 could improve the proliferative activity of spleen T cells and B cells and boost phagocytic activity of peritoneal macrophages at the concentration of 50 μg/ml (p < 0.05,p < 0.01). [ABSTRACT FROM PUBLISHER]

To investigate the association between the metalloproteinase-9 (MMP9) −1562C/T polymorphism and diabetic nephropathy (DN) in Han Chinese, the patients with type 2 diabetes were collected and divided into the non-DN (NDN) and DN groups; controls were recruited. Genotype and allele frequencies were assessed using polymerase chain reaction and restriction fragment length polymorphism. Results showed that SBP, DBP, HbA1c, UAER, Cr, BUN, TG, and TC were higher in the DN group compared with the control and NDN groups. SBP, HbA1c, and TC in DN patients with the TT and CT genotypes were lower than in those with CC. Compared with controls, the frequency of the T allele in the DN group was significantly lower. The MMP9 −1562C allele, SBP, Cr, BUN, TG, and TC were independent risk factors for DN. All of the above suggested that the MMP9 −1562C/T polymorphism was associated with DN in Han Chinese. [ABSTRACT FROM AUTHOR]

Aim The aim of this study was to investigate whether a functional insertion/deletion polymorphism (rs3783553) located in the 3′-untranslated regions of interleukin-1 A is associated with susceptibility to endometrial cancer (EC) in Chinese Han women. Methods A total of 272 patients and 294 age-matched healthy controls were included in the study. The genotype of the rs3783553 polymorphism was determined by polymerase chain reaction polyacrylamide gel electrophoresis. Results Our results showed that a significantly reduced risk of EC was found to be associated with the II (insertion/insertion) genotype compared with DD (deletion/deletion) only ( P = 0.037, odds ratio = 0.56, 95% confidence interval = 0.32-0.97) as well as compared with the DD + DI (deletion/insertion) genotype ( P = 0.032, odds ratio = 0.57, 95% confidence interval = 0.34-0.96) in the recessive model. No significant association was detected between certain clinical features of EC patients and genotype of rs3783553. Conclusions This study demonstrates that the rs3783553 polymorphism may be involved in susceptibility to EC. The II genotype seems to be a protective factor for EC in Chinese Han women. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas aeruginosa strains that are classed as extensively drug resistant (XDR-PA) are resistant to all antibiotics except for one or two classes and are frequently the cause of hard-to-treat infections worldwide. Our study aimed to characterize clinical XDR-PA isolates recovered during 2011-2012 at nine hospitals in the Hunan province of China. Methods: Thirty-seven non-repetitive XDR-PA strains from 37 patients were investigated for genes encoding antimicrobial resistance determinants, efflux pumps, outer membrane proteins, and movable genetic elements using polymerase chain reaction (PCR). The expression of genes encoding the efflux pump component MexA and the outer membrane protein OprD was measured using real-time PCR. In addition, clonal relatedness of these XDR-PA isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Results: Various genes encoding antimicrobial resistance determinants were found in all isolates. In particular, the blaTEM-1' blaCARB, armA, blaIMP-4' blaVIM-2' and rmtB, were found in 100, 37.8, 22, 22, 19 and 5 % of the isolates, respectively. Remarkably, two isolates coharbored blaIMP-4' blaVIM-2' and armA. In all 37 antibiotic-resistant strains, the relative expression of oprD was decreased while mexA was increased compared to the expression of these genes in antibioticsusceptible P. aeruginosa strains. All of the XDR-PA isolates harbored class I integrons as well as multiple other mobile genetic elements, such as tnpU, tnp513, tnpA (Tn21), and merA. A high genotypic diversity among the strains was detected by PFGE. Conclusions: Multiple antibiotic-resistance mechanisms contributed to the drug resistance of the XDR-PA isolates investigated in this study. Thus, the XDR-PA isolates in this area were not clonally related. Instead, multiple types of movable genetic elements were coharbored within each XDR-PA isolate, which may have aided the rapid development of these XDR-PA strains. This is the first report of XDR-PA strains that coharbor blaIMP-4' blaVIM-2' and armA. [ABSTRACT FROM AUTHOR]

Background: Serum preβ1-high density lipoprotein (preβ1-HDL) was defined by two-dimensional non-denaturing linear gel electrophoresis and apolipoprotein A-I immuno-blotting. Serum preβ1-HDL seems to play an important role in reverse cholesterol transport, a well-known anti-atherosclerosis process. However, there are still debatable questions for its quantification and coronary artery disease (CAD) relevance. Methods: We isolated the preβ1-HDL using a new native polyacrylamide gel electrophoresis (PAGE) system and lipid pre-staining serum. We established a two-demensional gel electrophoresis system. Results: We measured the preβ1-HDL in Tangier disease patients and subjects with cholesterol ester transfer protein (CETP) mutation. The preβ1-HDL is clearly separated from lipid-free apoA-I monomer and cannot be converted into other HDL particles under lecithin-cholesterol acyltransferase (LCAT) inhibition. This preβ1-HDL is a spheroidal particle with the highest apoA-1/cholesterol ratio and highest density (=1.21 g/ml), as compared with all other HDLs. Importantly, we found that serum from subjects with Tangier disease or with cholesterol ester transfer protein (CETP) mutation have no detectible preβ1-HDL particles. We recruited a total of 102 subjects underwent diagnostic coronary angiography and measured their preβ1-HDL levels. Among them, 56 had no stenosis of coronary artery and 46 were diagnosed as CAD, which was predefined as the presence of a luminal diameter stenosis =50 % in at least 1 major coronary artery territory. We found that preβ1-HDL is independently and negatively associated with the severity of the coronary artery stenosis (Gensini score). Conclusion: We established a novel and simple method for human serum preβ1-HDL quantification. We found β [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Dendrobium aurantiacum var. denneanumis widespread in southern China, locally known as “Shihu”, “Huangcao” or “Fengdou”, has long been used in traditional Chinese medicine for antipyretic, immunomodulatory, anti-aging effects and eye benefiting. Aim of this study To investigate the effects of gigantol extracted from the stem of D. aurantiacum var. denneanum on the formation of galactose-induced cataractogenesis and the potential mechanisms underlying these effects. Materials and methods Cataract lens models were induced by d -galactose both in vitro and in vivo . The transparency of the rat lenses in vitro and in vivo was observed with an anatomical microscope and a slit lamp microscope. The differential protein and action targets of gigantol were determined and compared among the control group, model group, and gigantol group using two-dimensional electrophoresis and mass spectrometry (MS). Enzyme kinetics was used to show the ability of gigantol to respress aldose reductase (AR) and inducible nitric oxide synthase (iNOS). Quantitative real-time PCR (RT-qPCR). was used to detect repression of the expression of AR and iNOS genes. Molecular docking and dynamic simulation were used to predict the interaction points and combination patterns between gigantol, AR, and iNOS. Results Gigantol was found to prevent galactose-induced damage to the rat lenses both in vitro and in vivo , to delay lens turbidity, and to keep the lenses transparent. Differential proteomes, MS, and RT-qPCR showed AR and iNOS to be the target proteins of gigantol. Gigantol reduced the galactose-induced AR and iNOS mRNA expression by 51.2% and 60.9%, respectively. The IC50 of gigantol for inhibition of AR and iNOS activities were 65.67 μg/mL and 8.768 μg/mL, respectively. Gigantol–AR binding sites were Trp111, His110, Tyr48, and Trp20, and gigantol–iNOS binding sites were Ile195 and Gln257. The main forms of interaction were hydrophobic forces, hydrogen bonds, and van der Waals forces. Conclusion Gigantol extracted from D. aurantiacum var. denneanum was found to inhibit galactose-induced formation of cataracts through repression of the gene expression and activity of AR and iNOS. [ABSTRACT FROM AUTHOR]

An increasing body of evidence has indicated that polymorphisms in the miRNA binding site of target gene can alter the ability of miRNAs to bind their target genes and modulate the risk of cancer. We aimed to investigate the association between a miR-520a binding site polymorphism rs141178472 in the PIK3CA 3′-UTR and the risk of colorectal cancer (CRC) in a Chinese Han population. The polymorphism rs141178472 was analyzed in a case-control study, including 386 CRC patients and 394 age- and sex-matched controls; the relationship between the polymorphism and the risk of colorectal cancer was examined. Individuals carrying the rs141178472 CC genotype or C allele had an increased risk of developing CRC (CC versus TT, OR (95% CI): 1.716 (1.084–2.716), P=0.022; C versus T, OR (95% CI): 1.258 (1.021–1.551), P=0.033). Furthermore, the expression of PIK3CA was detected in the peripheral blood mononucleated cell of CRC patients, suggesting that mRNA levels of PIK3CA might be associated with SNP rs141178472. These findings provide evidence that a miR-520a binding site polymorphism rs141178472 in the PIK3CA 3′-UTR may play a role in the etiology of CRC. [ABSTRACT FROM AUTHOR]

Objective: To study the effect of Tiantai No. 1 (天泰1号) on gene expression profile in hippocampus of Alzheimer's disease (AD) rat, molecular genetic target points of the effect of this drug were defined, its molecular genetic pharmacodynamic mechanism of anti-AD was further explored at molecular gene level, and a scientific basis was provided for its clinical availability and promotion. Methods: Thirty male Sprague-Dawley rats were divided into three groups with 10 rats per group: sham-operation group, model group and Tiantai No. 1 group. Sterile surgical procedure was applied, the model group with bilateral hippocampal injection of Aβ was established, and normal saline was used instead of Aβ in the sham-operation group. One week after the models was made, rats were administered by gastric lavage once every day for three consecutive weeks. The rats of the sham-operation group and the model group were daily fed with purified water by lavage; the rats of the Tiantai No.1 group treated group were administered with Tiantai No.1 by lavage. Total RNAs of hippocampus tissues were extracted with Trizol, the changes of hippocampus gene expression profiles in the above three groups were analyzed by using Affymetrix rat whole genome expression profile microarray. Results: Microarray analysis showed that, compared with the sham-operation group, the hippocampus of the model group had 50 up-regulated genes with significant difference (fold change >2), and 21 down-regulated genes with significant difference (fold change <0.5); compared with the hippocampus of the model group, the hippocampus of the Tiantai No. 1 group was found to have 5 up-regulated genes with significant difference (fold change >2) and 20 down-regulated genes with significant difference (fold change <0.5). The functions of differentially expressed genes of the groups were involved in nervous system's development, neuronic differentiation and function-regulation, cellular growth and differentiation and apoptosis, synaptic occurrence and plasticity, inflammation and immune response, ion channels/transporters, cellular signal transduction, cellular material/energy metabolism and so on. Conclusion: Tiantai No. 1 can regulate hippocampal function, and further regulate the brain function of animals in multiple gene target points by a number of ways. [ABSTRACT FROM AUTHOR]

Shigella flexneri is one of the major etiologic causes of shigellosis in Guizhou Province, China. However, the genetic characteristics of circulating isolates are unknown. Phenotypic and molecular profiles of 60 S. flexneri isolates recovered in Guizhou between 1972 to 1982 and 2008 to 2010 were determined. Nine serotypes (1a, 2a, 3a, 1b, 2b, X, Y, 4av and Yv) were identified. Multi-locus sequence typing differentiated the isolates into 20 sequence types (STs); 18 were novel. Four STs, ST 129, ST 100, ST 126 and ST 18, were most abundant, accounting for 65% of the isolates. Thirty-nine NotI-pulsed field gel electrophoresis patterns (pulsotypes, PTs) were observed; eight PTs were represented by more than one isolate with six isolates sharing the PT 13 profile. Multi-locus variable-nucleotide tandem-repeat analysis recognized 44 different types (MTs); seven MTs were represented by more than one isolate and MT 1 was most commonly encountered. Correlation between genetic relationships and serotypes was observed among the isolates studied; the majority of isolates belonging to the same serotype from different years clustered together based on the molecular data. These clustered isolates were also from similar geographical origins. These results enhance our understanding of genetic relationships between S. flexneri in Guizhou Province and can be used to help understand the changing etiology of shigellosis in China. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of Chaiqin Chengqi Decoction (柴芩承气汤,CQCQD) on cholecystokinin receptor 1 (CCKR1)-mediated signal transduction of pancreatic acinar cell in rats with acute necrotic pancreatitis (ANP). Methods: Twenty-seven Sprague-Dawley rats were randomized into three groups: the control group, the ANP group, and the CQCQD group (9 in each group). ANP rats were induced by two intraperitoneal injections of 8% L-arginine (pH=7.0, 4.4 g/kg) over a 2-h period. Rats were treated with 1.5 mL/100 g body weight of CQCQD (CQCQD group) or physiological saline (control and ANP groups) at 2 h interval. And 6 h after induction, pancreatic tissues were collected for histopathological examination. Pancreatic acinar cells were isolated for determination of CCKR1 mRNA and protein expression, phospholipase C (PLC) and inositol-1,4,5-triphosphate (IP3), and determination of fluorescence intensity (FI) as a measure of intracellular calcium ion concentration [Ca]. Results: The pancreatic histopathological score (6.2±1.1) and the levels of PLC (1,187.2±228.2 μg/mL) and IP3 (872.2±88.4 μg/mL) of acinar cells in the ANP group were higher than those in the control (2.8±0.4, 682.5±121.8 μg/mL, 518.4±115.8 μg/mL) and the CQCQD (3.8±0.8, 905.3±78.5 μg/mL, 611.0±42.5 μg/mL) groups ( P<0.05). [Ca] FI for the ANP group (34.8±27.0) was higher than that in the control (5.1±2.2) and CQCQD (12.6±2.5) groups ( P<0.05). The expression of pancreatic acinar cell CCKR1 mRNA in the ANP group was up-regulated (expression ratio=1.761; P=0.024) compared with the control group. The expression of pancreatic acinar cell CCKR1 mRNA in the CQCQD group was down-regulated (expression ratio=0.311; P=0.035) compared with the ANP group. The ratio of gray values of the CCKR1 and β-actin in the ANP group (1.43±0.17) was higher than those in the control (0.70±0.15) and CQCQD (0.79±0.11) groups ( P<0.05). Conclusions: Pancreatic acinar cell calcium overload of ANP induced by L-arginine was related to the up-regulated expressions of pancreatic acinar cell CCKR1 mRNA and protein. CQCQD can down-regulate expressions of pancreatic acinar cell CCKR1 mRNA and protein to reduce the PLC and IP3 of pancreatic acinar cells, relieving the calcium overload and reducing the pathological changes in rats with ANP. [ABSTRACT FROM AUTHOR]

Objective: To investigate the regulatory effects of Shenfu Injection (SFI, 参附注射液) on hemodynamic parameters and serum proteins in rats with post-infarction chronic heart failure (CHF). Methods: Forty-five healthy Wistar rats were randomized into three groups: sham, heart failure (model) and SFI group. The CHF was induced by left coronary artery ligation. Seven days after the surgical operation, animals in the sham group and the model group received saline (6.2 mL/kg/d), while animals in the SFI group received SFI (6.2 mL/kg d) intraperitoneally. Four weeks later, cardiac hemodynamic parameters were measured via the carotid route. The expression of serum proteins was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS). Results: Recording of hemodynamic parameters showed that left ventricular systolic pressure (LVSP), maximum rate of left ventricular pressure (+dp/dt) rise, and maximum rate of left ventricular pressure (−dp/dt) decrease, while the left ventricular end diastolic pressure (LVEDP) rose in the model group compared to those in the sham group ( P <0.05). The results of the MALDI-TOF MS indicated that haptoglobin (HP), pentraxin 3 (PTX3) and alpha-1-antitrypsin were up-regulated, while serum albumin and 40S ribosomal protein were down-regulated in the model group ( P <0.05). Compared with the model group, LVSP, +dp/dt and −dp/dt were higher, while LVEDP was lower in the SFI group ( P<0.05). Expression levels of HP and PTX3 were lower than in the model group ( P<0.05). Conclusion: SFI could improve hemodynamic function and decrease inflammatory reactions in the pathophysiology of CHF. The serum proteins HP and PTX3 could be potential biomarkers for chronic ischemic heart failure, and they could also be the serum protein targets of SFI. [ABSTRACT FROM AUTHOR]

Aim To explore the relationship between estrogen metabolism enzyme gene polymorphism and susceptibility to uterine fibroids, and to seek the screening molecular markers for genetic traits in uterine fibroid populations. Methods A total of 300 female Han Chinese patients and 300 healthy female Han Chinese volunteers in Nanjing (age range, 30-50 years) were recruited from Zhongda Hospital, Southeast University from February 2011 to March 2012. The single nucleotide polymorphisms ( SNP) of estrogen-metabolizing enzyme genes from the two groups of women were examined by polymerase chain reaction denaturing high-performance liquid chromatography, which were four COMT gene loci including rs3087869, rs165774, rs165599 and rs4680, three CYP1A1 gene loci including rs1048943, rs4646421 and rs4646422, and three CYP1B1 gene loci including rs1056827, rs1056836 and rs1056837. Genotype frequencies among cases and controls were calculated and analyzed by binary logistic regression. Results Regression analysis of SNP showed that COMT IVS1+2329 C>T (odds ratio [ OR], 2.872; 95% CI, 1.690-4.882) and Val158 Met ( OR, 2.593; 95% CI, 1.546-4.350), CYP1A1 Ile462 Val ( OR, 2.383; 95% CI, 1.418-4.005) and Gly45 Asp ( OR, 2.489; 95% CI, 1.49-4.159), and CYP1B1 Ala119 Ser ( OR, 3.361; 95% CI, 2.035-5.552) and Leu432 Val ( OR, 0.164; 95% CI, 0.061-0.441) influenced uterine fibroids significantly ( P < 0.05). Allele and genotype frequencies among cases and control were calculated and examined to match the Hardy-Weinberg equilibrium with the χ2-test. Conclusion The genetic polymorphisms of IVS1+2329 C>T and Val158 Met loci in COMT, Ile462 Val and Gly45 Asp loci in CYP1A1 and Ala119 Ser loci in CYP1B1 were risk factors for uterine leiomyoma development, and Leu432 Val locus in CYB1B1 may be a protective factor. The results provide a theoretical basis for genetic screening and early intervention for uterine leiomyoma-susceptible populations. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Chan Su, an ethanolic extract from skin and parotid venom glands of the Bufo bufo gargarizans Cantor, is widely used as a traditional Chinese medicine for cancer therapy. Although the anti-cancer properties of Chan Su have been investigated, no information exists regarding whether Chan Su has genotoxic effects in cancer cells. The aim of the present study was to examine the cyto-/genotoxic effect of Chan Su in human breast carcinoma (MCF-7 cells), human lung carcinoma (A-549 cells), human T cell leukemia (Jurkat T cells), and normal human lymphocytes. Materials and methods: Effects on the viability of MCF-7, A-549, Jurkat T cells, and normal lymphocytes were evaluated by Trypan blue exclusion assays. The DNA content in the sub-G1 region was detected by propidium iodide (PI) staining and flow cytometry. The genotoxicity of Chan Su was assessed by single-cell gel electrophoresis (comet assay) and the cytokinesis-block micronucleus assay (CBMN assay). Results: Chan Su significantly inhibited the viability of MCF-7, A-549, and Jurkat T cells dose dependently, but had no effect on normal human lymphocytes. Apoptotic death of the cancer cells was evident after treatment. Chan Su also induced genotoxicity in a dose-dependent manner, as indicated by the comet and cytokinesis-block micronucleus assays. Conclusions: These findings suggest that Chan Su can induce apoptotic death of, and exert genotoxic effects on, MCF-7, A-549, and Jurkat T cells. [Copyright &y& Elsevier]

The Xuefuzhuyu capsule (XFZY) is widely used for the treatment of ischemic heart disease (IHD) in China. We previously demonstrated that XFZY could reduce apoptosis in Sprague-Dawley rat cardiomyocytes with the similar effect of resveratrol (Res) Hori et al. (2013), although its molecular mechanism underlying this protective effect is still unclear. Silent information regulator of transcription 1 (SIRT1) had been demonstrated to be responsible for cardioprotection against ischemia-reperfusion injury via long-term transcriptionally regulatory mechanism Braunersreuther and Jaquet (2012). Therefore, in the present study, we aimed to test if XFZY might contribute to the protection of ischemic myocardial cells induced by ischemia through SIRT1-mediated signal transduction pathway by using electron micrograph, RT-PCR assay, and western-blot test. All the result showed that the target genes of SIRT1 pathway including P53, NF-kB, FOXO1, FOXO3, and FOXO4 were significantly downregulated to SIRT1, suggesting that apoptosis pathway might transcriptionally be regulated to SIRT1. In addition, the expression level of SIRT1 was significantly increased by XFZ, it might prove that SIRT1 is the target of XFZY working on ischemia heart disease. Our findings supported that XFZY might function to protect myocardial cells and reduce myocardial injury though SIRT1 signaling pathway and has the same pharmacological effect with Res. [ABSTRACT FROM AUTHOR]

Abstract: A total of 152 Salmonella isolates recovered from retail foods in Henan Province in China were characterized using serotyping and antimicrobial susceptibility testing. Nalidixic acid and/or ciprofloxacin-resistant isolates were further examined for amino acid substitution of gyrase and topoisomerase Ⅳ, and for the presence of qnr (qnrA, qnrB and qnrS) and aac(6′)-Ib genes. Selected serovars were subtyped using pulse field gel electrophoresis (PFGE). Among all isolates tested, Salmonella Enteritidis (34.2%) was the most common serotype detected, followed by S. Indiana (9.9%), S. Derby (9.9%), S. Agona (6.6%), S. Typhimurium (5.3%) and S. Albany (4.6%). Twenty-eight percent of the isolates were resistant to 1–3 antimicrobials, 37% to 4–6 antimicrobials, 23% to 7–9 antimicrobials, and 12% to more than 10 antimicrobials. Resistance was most frequently detected to sulfamethoxazole (95.3%), followed by trimethoprim/sulfamethoxazole (80.5%), tetracycline (75.8%), nalidixic acid (75.8%), ampicillin (45.6%), chloramphenicol (35.6%), streptomycin (32.9%), kanamycin (24.8%), gentamicin (21.5%) and amoxicillin/clavulanic acid (14.8%). Resistance was also observed to ciprofloxacin (12.1%), cefoperazone (18.8%), ceftriaxone (6.0%) and cefoxitin (4.0%). A total of 45 amino acid substitutions were identified in quinolone resistant determination region (QRDR) of gyrA and parC of 19 ciprofloxacin-resistant isolates. The most common mutations in gyrA were Ser83Phe and Asp87Gly, in parC was Ser80Arg. QnrA, qnrB, qnrS and aac(6′)-Ib genes were identified in 55 (46.6%), 15 (12.7%), 23 (19.5%) and 16 (13.6%) of 118 nalidixic acid resistant but ciprofloxacin susceptible or less resistant isolates, respectively. A total of 108 PFGE patterns were generated among 146 selected isolates. Our findings indicate that foodborne Salmonella isolates in Henan, China were phenotypically and genotypically diverse and many isolates demonstrated multidrug resistance. [Copyright &y& Elsevier]

Background: We compared the T cell antigen receptor (TCR-BV) gene families of peripheral blood mononuclear cells (PMBC) between children with tuberculosis (TB) and those inoculated with the Bacille Calmette Guerin (BCG) vaccine. Methods: The total RNA was extracted from PMBC of 15 TB children, 15 BCG-vaccinated children and 15 healthy controls. The RNAs were reverse-transcribed and amplified by polymerase chain reaction (PCR). PCR products were separated on 1.5% agarose gel and analyzed with the Genescan technique. Results: Some TCR-BV gene families in TB children and BCG-vaccinated children exhibited a blur band in the predicted position on 1.5% agarose gel, some showed a distinct or fainted band. In general, many shared predominant clonal TCR-BV gene families (Vß2, Vß16, Vß21, Vß22) and the restricted-expression families (Vβ14 and Vβ17). All the gene families of the control children only exhibited blur bands and polyclonal. Conclusions: The skewed profile of TCR-BV gene families in TB children and BCG-vaccinated children are similar, which may probably explain the protective effects of BCG-vaccine against TB in children. [ABSTRACT FROM AUTHOR]

Background: In Traditional Chinese Medicine (TCM), tongue diagnosis has been an important diagnostic method for the last 3000 years. Tongue diagnosis is a non-invasive, simple and valuable diagnostic tool. TCM treats the tongue coating on a very sensitive scale that reflects physiological and pathological changes in the organs, especially the spleen and stomach. Tongue coating can diagnose disease severity and determine the TCM syndrome ("Zheng" in Chinese). The biological bases of different tongue coating appearances are still poorly understood and lack systematic investigation at the molecular level. Methods: Tongue coating samples were collected from 70 chronic gastritis patients and 20 normal controls. 16S rRNA denatured gradient gel electrophoresis (16S rRNA-DGGE) and liquid chromatography and mass spectrometry (LC-MS) were designed to profile tongue coatings. The statistical techniques used were principal component analysis and partial least squares-discriminate analysis. Results: Ten potential metabolites or markers were found in chronic gastritis patients, including UDP-D-galactose, 3-ketolactose, and vitamin D2, based on LC-MS. Eight significantly different strips were observed in samples from chronic gastritis patients based on 16S rRNA-DGGE. Two strips, Strips 8 and 10, were selected for gene sequencing. Strip 10 sequencing showed a 100% similarity to Rothia mucilaginosa. Strip 8 sequencing showed a 96.2% similarity to Moraxella catarrhalis. Conclusions: Changes in glucose metabolism could possibly form the basis of tongue coating conformation in chronic gastritis patients. The study revealed important connections between metabolic components, microecological components and tongue coating in chronic gastritis patients. Compared with other diagnostic regimens, such as blood tests or tissue biopsies, tongue coating is more amenable to, and more convenient for, both patients and doctors. [ABSTRACT FROM AUTHOR]

Abstract: For centuries, Patrinia heterophylla had been used in China to treat many diseases including tumor. Triterpenes has been identified as the major active constituents in Patrinia heterophylla. To elucidate the antitumor mechanism of triterpenes from Patrinia heterophylla1 (TPH), a proteomic analysis is carried out with TPH treatment in K562 cells. The total proteins extracted from TPH treated K562 cells are analyzed by two dimensional gel electrophoresis (2-DE) and compared with those untreated K562 cells. Mass spectrometry is applied to identify the differentially expressed proteins. Twenty-three differentially expressed significant proteins are discovered. Eight proteins are later identified by mass spectrometry (MALDI-TOF-MS) and Mascot software. Among them, four proteins are up-regulated (Aldolase A, Glyceraldehyde-3-phosphate dehydrogenase, Flavin reductase and Hemoglobin subunit) and four proteins were down-regulated (Heat-shock protein 90 〈Alpha〉 (HSP90-〈Alpha〉), Eukaryotic translation initiation factor 5A, Moesin, tublin) by TPH treatment in K562 cells. The identified proteins are associated with energy metabolism, oxidative stress, apoptosis, signal transduction, differential induction, and protein biosynthesis. These findings might provide valuable insights into the antitumor mechanism of TPH in K562 cells. [Copyright &y& Elsevier]

Aims Hepatocyte nuclear factor-1α (HNF-1α) regulates the expression of genes encoding proteins involved in glucose metabolism and insulin secretion. Mutations in the HNF-1α gene cause maturity-onset diabetes of the young Type 3. However, the mechanism leading to this disease has not been completely ascertained. Previously, we found a novel mutation in the regulatory element of the human HNF-1α gene in two Chinese diabetes pedigrees. The nucleotide at position -128 T was substituted by G (nt-128 T→G). In this study, we analysed the functional defect of nt-128 T→G in HNF-1α transcription activity. Methods Luciferase reporter gene assays were carried out to examine the functional characteristics of this mutant. Electrophoretic mobility shift assays and chromatin immunoprecipitation were performed to confirm the binding of nuclear proteins to oligonucleotides. Results The variant construct (nt-128 T→G) had a 1.65-fold increase in promoter activity compared with that of the wild-type construct in HepG2 cells and a 1.33-fold increase in MIN6 cells, respectively. The variant resided at a FOXA/HNF-3 binding site identified by a series of competitive electrophoretic mobility shift assays and antibody supershift analyses. The assays showed a differential binding affinity in the wild-type and the nt-128 T→G mutant fragments by FOXA/HNF-3. Chromatin immunoprecipitation indicated that FOXA/HNF-3 bound to this region in vivo. One nucleotide substitution in the FOXA/HNF-3 site in the human HNF-1α regulatory element caused an increase of HNF-1α transcriptional activity. Conclusions Our data suggested that this substitution in the promoter region affects DNA-protein interaction and HNF-1α gene transcription. The mutant may contribute to the development of diabetes in these two nt-128 T→G pedigrees of Chinese. [ABSTRACT FROM AUTHOR]

Coxsackievirus B3(CVB3) infection is the major cause of viral myocarditis, as well as dilated cardiomypathy. Rhubarb is one of the oldest and best-known traditional Chinese medicines. We initiated this study to determine the antiviral effect of an ethanol extract from the roots and rhizoma of Rheum palmatum ( R. palmatum, one of the Chinese Rhubarbs), against CVB3 in tissue culture cells and in a mouse model. The ethanol extract from R. palmatum showed significant inhibitory activity against CVB3 on HEp-2 cells when added after infection, with IC50 of 4 μg/ml, TI of 10. The medicated mouse serum still contained the pharmaceutical compound 24 h after intraperitoneal injection, and exhibited an antiviral effect on CVB3-infected cells, especially in the 0.3 and 0.5 g/kg/day treatment groups. Furthermore, the CVB3-infected mice were treated with the extract solution with dosages of 0.3 g/kg/day beginning 24 h post-CVB3 exposures. The ethanol extract treated mice showed alleviated clinical signs, better survival rate, prolonged MTD and decreased viral titers compared to the virus control group. Our results indicate that the ethanol extract from R. palmatum has the anti-CVB3 activity in vitro and in vivo and thus provides a re-evaluation of this old remedy with a broad therapeutic potential. [ABSTRACT FROM AUTHOR]

Ganoderma lucidum (Fr.) Karst is a traditional Chinese herb that has been widely used for centuries to treat various diseases including cancer. Herein, an ethanol-soluble and acidic component (ESAC), which mainly contains triterpenes, was prepared from G. lucidum and its anti-tumor effects in vitro were tested on human breast cancer cells. Our results showed that ESAC reduced the cell viability of MCF-7 and MDA-MB-231 cells in a concentration-dependent manner with IC50 of about 100 μg/mL and 60 μg/mL, respectively. DNA damage was detected by Comet assay and the increased expression of γ-H2AX after ESAC treatment was determined in MCF-7 cells. Moreover, ESAC effectively mediated G1 cell cycle arrest in both concentration- and time-dependent manners and induced apoptosis as determined by Hoechst staining, DNA fragment assay and Western blot analysis in MCF-7 cells. In conclusion, ESAC exerts anti-proliferation effects by inducing DNA damage, G1 cell cycle arrest and apoptosis in human breast cancer cells. [ABSTRACT FROM AUTHOR]

Abstract: A fucosylated polysaccharide sulfate, AMP-2, was purified by DEAE-Sepharose Fast Flow and Sephadex G-100 columns in successive steps from a special sea cucumber in southeastern China. HPLC and cellulose acetate membrane electrophoresis experiments confirmed AMP-2 was a homogenous carbohydrate with a relative molecular weight of ca. 2.4×104 Da, and methylation analysis indicated that polysaccharide was composed of 1-substituted-Galp, 1,4-disubstituted-GalNp, 1,2-disubstituted-FucSp, 1,4,6-trisubstituted-Glcp in a molar ratio of ca. 0.5:2.0:1.0:3.0, together with a small amount of different substituted Manp. Sulfated derivative and carboxymethylated derivative were prepared using dry pyridine and chlorosulfonic acid, and chloroacetic acid, respectively. Anticoagulant activities in vitro investigation showed that sulfated derivative showed a stronger ability than native polysaccharide and carboxymethylated derivative, which might be caused by their different percentages and types of functional groups in their structures. [Copyright &y& Elsevier]

Abstract: Beta-Hemolytic group G streptococci cause a considerable invasive disease burden and sometimes disease outbreaks. Little is known about the critical epidemiologic parameter of genetic relatedness between isolates. We determined the emm types of 65 Streptococcus dysgalactiae subsp. equisimilis isolates. We formulated multilocus sequence typing (MLST) primers with 6 of the 7 loci corresponding to the Streptococcus pyogenes MLST scheme. We performed MLST with 69 S. dysgalactiae subsp. equisimilis isolates to represent each emm type identified. These strains were further analyzed by pulsed-field gel electrophoresis (PFGE) typing. Sixteen emm types were observed. Eighteen unique combinations of allelic profiles (sequence types [STs]) were obtained with 12 profiles each accounting for multiple isolates. Forty-one MLST STs were observed. Analysis of the PFGE patterns generated revealed 10 clones. Over 80% of the isolates were distributed in 3 large clones. Isolates within 16 redundantly represented S. dysgalactiae subsp. equisimilis emm types shared identical or nearly identical STs and subtypes of PFGE, demonstrating concordance between the emm type and genetic relatedness. It is conceivable that some particular characteristics in the genomes of these strains are responsible for their predominance in different regions. [Copyright &y& Elsevier]

The article presents a study on the prevalence and the phylogeny of leishmaniasis in canines in Beichuan County, Sichuan, China. The study used several methods including enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) and multilocus enzyme electrophoresis (MLEE). Results show that 13 among the 86 dogs were found positive for the disease using the dip-stick test. It concludes that more control is needed for the management of the disease since it poses a great public concern.

Objective. Primary OA and Kashin–Beck disease (KBD) show similar pathological changes in articular cartilage. The objective was to screen differentially expressed genes between OA and normal cartilage, confirm the candidate gene expression among OA, KBD and normal cartilage, and then clarify its role in vitro.Methods. Differentially expressed genes in OA cartilage were screened by suppression subtractive hybridization (SSH) and verified by real-time quantitative PCR (Q-PCR) analysis. Heparan sulphate 6-O-sulphotransferase 2 (HS6ST2) expression was identified by Q-PCR and immunohistochemistry. After suppressing HS6ST2 by RNA interference in C28/I2 human chondrocyte line, the effects were analysed through determining the cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), the aggrecan contents by toluidine blue staining and the mRNA expression levels of SRY-type high mobility group box 9 (SOX9), AGC1, MMP3, a disintegrin and metalloproteinase domain with thrombospondin motifs 4 (ADAMTS4) and ADAMTS5 by Q-PCR.Results. HS6ST2 in the reverse subtraction library was identified as a down-regulated gene in OA and KBD at both mRNA and protein levels. The percentage of safranion O staining area was correlated positively with the percentage of HS6ST2-positive chondrocytes in OA and KBD cartilage. After HS6ST2-specific short interfering RNA (siRNA) transfection to C28/I2 cells, the cell viability was inhibited significantly, and the mRNA expression levels of SOX9 and AGC1 were reduced markedly, while MMP3 expression was increased significantly.Conclusion. HS6ST2 down-regulation was identified in both OA and KBD cartilage. The findings first suggest that HS6ST2 may participate in the pathogenesis of OA and KBD by influencing aggrecan metabolism. [ABSTRACT FROM PUBLISHER]

It is widely accepted that biodiversity is lower in more extreme environments. In this study, we sought to determine whether this trend, well documented for macroorganisms, also holds at the microbial level for bacteria. We used denaturing gradient gel electrophoresis (DGGE) with phylum-specific primers to quantify the taxon richness (i.e., the DGGE band numbers) of the bacterioplankton communities of 32 pristine Tibetan lakes that represent a broad salinity range (freshwater to hypersaline). For the lakes investigated, salinity was found to be the environmental variable with the strongest influence on the bacterial community composition. We found that the bacterial taxon richness in freshwater habitats increased with increasing salinity up to a value of 1‰. In saline systems (systems with >1‰ salinity), the expected decrease of taxon richness along a gradient of further increasing salinity was not observed. These patterns were consistently observed for two sets of samples taken in two different years. A comparison of 16S rRNA gene clone libraries revealed that the bacterial community of the lake with the highest salinity was characterized by a higher recent accelerated diversification than the community of a freshwater lake, whereas the phylogenetic diversity in the hypersaline lake was lower than that in the freshwater lake. These results suggest that different evolutionary forces may act on bacterial populations in freshwater and hypersaline lakes on the Tibetan Plateau, potentially resulting in different community structures and diversity patterns. [ABSTRACT FROM AUTHOR]

Aims: The aim of this study was to characterize the different structures of microbial communities between 20 healthy women and 17 bacterial vaginosis (BV)-positive women of reproductive age using denaturing gradient gel electrophoresis (DGGE). Material and Methods: Vaginal samples from 17 BV-positive and 20 BV-negative women were subjected to DNA extraction, and amplified with eubacterial 16S rRNA gene-specific primers via polymerase chain reaction. The polymerase chain reaction products were separated using DGGE. Bands were excised, re-amplified, purified and sequenced. DNA sequences were compared with GenBank database. Phylip software packages were used to calculate sequencing data and form a phylogenetic tree to identify the genetic relations for microbiota inhabited in vaginal ecosystems of BV-positive women. Results: In total, 28 kinds of organisms were detected that comprised BV(+) vagina microbial community, varying from three to nine kinds with an average of 5.71 kinds per woman. Only seven species were detected in BV(−) women, ranging between one and five species with an average of 2.40 species per woman, which was significantly lower than that detected in BV(+) women ( t = 7.39, P < 0.001). A strain of Uncultured Lactobacillus sp. clone EHFS1_S05c (29/37; 78.38%) was most commonly presented in both BV-negative and BV-positive women, but the mean proportion of this Lactobacillus sp. strain to the whole microbial population colonized in the vaginal tract of BV(−) women was sharply higher than that calculated from BV(+) women ( t = 2.92, P < 0.01). Conclusions: The findings indicate further diversity in the category of vaginal microorganisms associated with BV. The presence of Gardnerella vaginalis is not necessary as a sign for gynecologists to determine whether or not a woman is affected by BV. [ABSTRACT FROM AUTHOR]

Background: protein glycosylation varies with the physiological and pathological status of the cell. Consequently, analysis of protein-linked glycans has growing importance both in basic glycobiological research and as a potential tool for monitoring the physiological state in humans.Design, setting and participants: a total of 265 healthy northern Chinese men and women were grouped by age and gender. The mean age in males and females was similar.Objective: the study is aimed to evaluate the effects of the age and gender on the human serum N-glycans profiles in the clinical diagnose of ageing and disease.Methods: the 265 human serum N-glycan profiles were obtained by DNA sequencer-assisted fluorophore-assisted carbohydrate electrophoresis. Comparison of N-glycan profiles was carried out among the different genders and age groups and the data were analysed with the GeneMapper software.Results: age-related changes in the three N-glycan structures (NGA2F, NGA2FB and NA2F) were observed. Interestingly, fucosylation of N-glycans was significantly different (P < 0.0001) between men and women: more core-α-1,6-fucosylated glycans were detected in women, whereas more branching-α-1,3-fucosylated N-glycans were seen in men.Conclusions: the N-glycome profile in serum is gender and age dependent. This should be taken into consideration in the development of serum glycome markers. [ABSTRACT FROM AUTHOR]

In order to detect the DNA from highly processed medicinal donkey-hide glue Colla Corii Asini from adulterants blended or totally substituted with horse, cattle, and pig tissues, various species-specific PCR assays based on highly repetitive SINE elements were established. For horse and donkey tissues, a two-step PCR assay was adopted, including an initial detection of equine DNA based on the equine SINE ERE-1 and a further detection of horse DNA based on the horse specific satellite. The assay can detect the equine DNA and the horse DNA from binary solid glue mixtures spiked with 0.1% Colla Corii Asini and 1% horse-hide glue respectively. In addition, for bovine and porcine detection, the primer pairs were designed upon 1.71 IB bovine repeat and PRE-1 respectively. The developed methods can detect bovine and porcine DNA from binary solid glue mixtures spiked with 0.1% cattle-hide glue and 0.1% pig-hide glue respectively. The result shows that our developed PCR assays were useful, convenient and sensitive for Colla Corii Asini identification. [ABSTRACT FROM AUTHOR]

Analysis of variable number tandem repeats (VNTRs) by polymerase chain reaction (PCR) is a common method used to predict engraftment status in post-allogeneic haematopoeitic stem cell transplantation (HSCT) patients. Different populations have different copies of repeated DNA sequence and hence, different percentage of informativeness between patient and donor. PCR amplification of four highly polymorphic VNTR markers (YNZ-22, D1S80, D1S111 and ApoB) was conducted on 60 patient-donor pairs. The informativeness of the markers was analysed using 3% agarose gel electrophoresis. We developed an algorithm for identification of informative VNTR markers in 60 post-allogeneic HSCT patients. YNZ-22 was the most informative (72%), followed by D1S80 (63%) and D1S111 (60%), while the least informative was ApoB (47%). The degree of informativeness achieved was 95%, which could discriminate 57 patient-donor pairs, when all four markers were combined. Since population genetic studies on VNTR loci are not well established in Southeast Asia, the present study is useful to determine reliable markers during the initial screening step of chimerism analysis. By following this algorithm, we are able to reduce time and cost of finding a suitable VNTR marker in our cohort. [ABSTRACT FROM AUTHOR]

Background: The community could be a reservoir of antibiotic resistance genes. The aim of this study was to investigate the prevalence and genetic environments of blaCTX-M among faecal Escherichia coli obtained from healthy persons in a region of China. Methods: Bacteria in stool specimens were screened for extended-spectrum ββ-lactamase (ESBL) production on 2 MacConkey agars, one with cefotaxime and one with ceftazidime. blaCTX-M and their genetic environments, as well as phylogenetic analysis and detection of the O25b-ST131 clone of E. coli, were characterized by polymerase chain reaction and sequencing. Pulsed field gel electrophoresis and conjugation assays were performed by standard procedures. Results: A surprisingly high number (50.5%, 55/109) of faecal samples showed the presence of ESBL-producing E. coli. blaCTX-M genes were detected in all of these strains. The CTX-M-9 group (41/55, 74.5%) was found most frequently, followed by the CTX-M-1 group (16/55, 29.1%). CTX-M-14 ( n == 39) was the predominant CTX-M enzyme in this study. However, the genes for the CTX-M-2 and CTX-M-8 groups were not observed. IS Ecp1 was detected in 90.9% of the strains, while IS 26 was observed upstream from blaCTX-M in only 1 strain. Phylogenetic groups A and D were found to predominate in commensal E. coli. High clonal diversity was observed and most blaCTX-M genes were transferable. The O25b-ST131 clone was found in 4 strains. Conclusions: This study reveals the wide dissemination of CTX-M ESBL-producing E. coli in the gut flora of healthy individuals in China. [ABSTRACT FROM AUTHOR]