OBJECTIVETo study the effect of bifidobacterium genomic DNA on umbilical cord blood mononuclear cell (CBMC), and investigate the immunoregulation of bifidobacterium DNA and explore possible mechanisms by which bifidobacterium acts against allergic reaction.

METHODSBifidobacterium genomic DNA (bDNA) and human DNA (hDNA) were extracted with phenol/chloroform/isoamyl alcohol and stored at -20 degrees C for later use. Parts of bDNA were completely digested with DNaseI (d-bDNA) at 37 degrees C. CBMCs were isolated with Ficoll from umbilical cord blood and incubated at 37 degrees C in a 5% CO2 humidified incubator. These cells were divided into four groups, control group: without any stimulant; bDNA group: stimulated with 25 microg/ml bDNA; d-bDNA group: stimulated with 25 microg/ml d-bDNA; hDNA group: stimulated with 25 microg/ml hDNA. The cells were stimulated with different stimulants in vitro, at the end of incubation culture supernatant and cells were collected. IL-12 and IL-10 levels in the culture supernatant were measured by enzyme linked immuno sorbent assay (ELISA); cells secreting IL-4 and IFN-gamma were counted by enzyme linked immunospot (ELISPOT) assay; and total RNA was isolated from the cells to assay T-bet and GATA3 mRNA expression levels by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSSix hours after stimulation there was no significant difference in IL-12 level in supernatant among the four groups; 12 hours after stimulation, IL-12 level in supernatant of bDNA treated group was significantly higher than that of each of the other groups, so were the results obtained at 24 hours and 48 hours after stimulation (P < 0.05). No significant difference could be detected in IL-12 level in supernatant among the other 3 groups. On the other hand, 6 hours after stimulation there was no significant difference in IL-10 level in supernatant among the four groups. But 12 and 24 hours after stimulation IL-10 level in supernatant of bDNA treated group was lower than that of each of the other groups, but the difference was not statistically significant. The count of IFN-gamma secreting cells of bDNA treated group was higher than that of the other groups, while IL-4 secteting cells of bDNA treated group were lower than that of the other groups. After bDNA stimulation, nuclear factor T-box expressed in T cells (T-bet) mRNA expression was conspicuously enhanced as compared to the other three groups (P < 0.05). GATA3 mRNA transcription in CBMC had no significant change after bDNA stimulation.

CONCLUSIONbDNA could promote secretion of Th1 type cytokine IL-12, while Th2 type cytokine IL-10 level of cell supernatant was decreased. bDNA could stimulate secretion of IFN-gamma by CBMC and inhibit secretion of IL-4. T-bet mRNA expression was highly enhanced after bDNA stimulation. bDNA could upregulate Th1 type response, which may be one of important mechanisms by which bifidobacterium inhibit allergic response.

Bifidobacterium ; cytology ; genetics ; Cell Culture Techniques ; DNA, Bacterial ; biosynthesis ; metabolism ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; immunology ; GATA3 Transcription Factor ; genetics ; Humans ; Infant, Newborn ; Interferon-gamma ; immunology ; secretion ; Interleukin-10 ; immunology ; secretion ; Interleukin-12 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Leukocytes, Mononuclear ; immunology ; secretion ; RNA, Messenger ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; T-Box Domain Proteins ; genetics ; Th1 Cells ; drug effects ; immunology ; secretion

Background The rapid diagnosis can win more treating opportunities for patients with fungal keratitis.Even though the fungal culture is the gold standard for the diagnosis of fungal keratitis,it is difficult in early diagnosis due to the long duration of cultivation and false-negative rate.Objective This trial was to explore the clinical value in the rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy.Methods Corneal scraping and laser scanning confocal microscopy were separately performed in 167 eyes of 167 patients with fungal keratitis.All the eyes were examined by the slit lamp,followed by laser scanning confocal microscope,and then the 10％ KOH corneal smear was examined under the optical microscope.Results The positive rate of diagnosis was 75％ (125/167) by corneal scraping,and that by laser scanning confocal microscopy was 91％ (152/167).The positive rate of examining outcome was significantly higher in laser scanning confocal microscopy than that of corneal scraping (x2 =14.88,P =0.00).The positive results were 114 cases and negative results were 4 cases by two methods,with the concordance rate 70.7％ (118/167).The hyphae or spore were seen in 32 cases by laser scanning confocal microscopy in 42 negative cases by corneal scraping,and in 15 negative cases by confocal laser scanning microscopy,11 positive outcomes were offered by corneal scraping.Conclusions The combined application of corneal scraping with confocal laser scanning microscopy can improve and speed up the diagnosis positive rate of fungal keratitis.

Objective To investigate the causes,consequences and management of injuries to the draining veins after embolization of brain arteriovenous malformations(BAVMs)with ?-n-butyl cyanoacrylate (NBCA).Methods The angiographic imaging data of 189 BAVMs patients who underwent NBCA embolization were studied retrospectively.The status of the draining veins before and after NBCA embolization was observed and compared.The intracerebral hemorrhage(ICH)complications and their relation to their angiographic features were analyzed.Results Twenty-three patients out of 189 patients showed injuries to the draining venous system,including 10 low-grade injury,6 moderate injury,and 7 high- grade injury.Six patients suffered from ICH after embolization,of whom 4 patients were due to injuries of the draining veins(2 moderate and 2 high-grade).In the 3 months follow-up evaluation of 4 patients with ICH, one died,one was in vegetative state,and the other two patients suffered from residual severe or minor (1 patient for each)permanent neurological deficits.Conclusion Our findings suggest that injury of the draining veins is the major cause of ICH and may lead to serious consequences after embolization of BAVMs with NBCA.

OBJECTIVETo investigate the risk factors for the prognosis in patients with gastric cancer undergoing surgery.

METHODSClinical data of 1031 cases who underwent gastric cancer resection from January 2003 to December 2007 were studied using univariable analysis and multivariable regression analysis.

RESULTSIn 1031 cases,95 (9.2%) cases were early-stage gastric cancer. The other 936 (90.8%) cases were advanced gastric cancer. The tumor was resectable in 980 (95.1%) cases, of which 874 (84.8%) were curative resection,106 (10.3%) were palliative, and 51 (4.9%) were bypass procedures or laparotomy alone. The stage-specific 5-year survival rates were 93.2% (stage IA), 65.1%(stage IB), 52.3% (stage II), 41.4% (stage IIIA), 16.6% (stage IIIB) and 10.6% (stage IV), respectively. The 1-, 3- and 5-year survival rates were 80.2%, 58.0% and 48.2%, respectively. The independent risk factors associated with the prognosis of these patients were tumor size, serum albumin, curative resection, TNM staging and multidisciplinary treatment in both univariable and multivariable analyses.

CONCLUSIONSEarly curative resection is the most important treatment for the patients with gastric cancer. Individualized surgical procedure combined with multidisciplinary treatment can improve the outcome. Tumor size, serum albumin level and TNM staging are important predictors of survival in patients with gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Causality ; Female ; Gastrectomy ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Stomach Neoplasms ; epidemiology ; mortality ; surgery ; Survival Rate ; Treatment Outcome

Since late 2010, porcine epidemic diarrhea virus (PEDV) has been re-emerging in central China. To explore the possible reason of the PEDV outbreaks, twelve PEDV field strains were isolated from different swine breeding farms in central China during 2010-2012, and molecular diversity, phylogenetic relationships of these strains with other PEDV reference strains were investigated. Sequence analysis of S, M and ORE3 genes revealed that the central China PEDV isolates had several specific nucleotides and amino acids which were different from PEDV reference strains. In addition, the entire S genes of eleven central China PEDV isolates were found to be nine nucleotides longer in length than CV777 and large number of amino acid variations was accumulated in the N-terminal region of S gene. Phylogenetic analysis showed that the central China PEDV isolates had close relationship with Korea strains (2007-2009), Thailand strains (2007-2008), Vietnam strains (2009-2010), Japan strains (2010), and other prevailing strains from other parts of China (2010-2012). However, they differed genetically from European strains (CV777, Brl/87), China strains (2003-2007) and the vaccine strains (CV777) used in China. These results imply that a rapid variation and evolution of central China PEDV strains has occurred in recent years, and a more efficient vaccine strain should be selected to prevent and control outbreaks of PEDV in China.
Animals ; China ; epidemiology ; Disease Outbreaks ; Feces ; virology ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Porcine epidemic diarrhea virus ; classification ; genetics ; isolation & purification ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; genetics

OBJECTIVETo study the chemical constituents of the dried roots of Zanthoxylum dimorphophyllumr. spinifolium and to find out the active constituents of the plant.

METHODModern chromatography was used to purify chemical constituents, and their structures were identified by various spectral methods.

RESULTFour compounds were isolated and identified as isopimpinellin (I), xanthoxyletin (II), 6-(2', 3'-dihydroxy-3'-methyl-butyl)-7-hydroxy-2H-1-benzopyran-2-one (III), 6-(2'-O-beta-D-glucopyranosyloxy, 3'-dihydroxy-3'-methybutyl)-7-hydroxy-2H-1-benzopyran-2-one (IV).

CONCLUSIONAll of the above compounds were isolated from the above mentioned plant for the first time.

Coumarins ; chemistry ; isolation & purification ; Furocoumarins ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Zanthoxylum ; chemistry

Objective To detect the resistance index and esterase activity of each generation of DDVP-resistant Culex mosquitoes and analyze the relationship between insecticide resistance and esterase. Methods WHO bioassay and micro-plate measurement were used for the detection. Results The resistance index increased to 12.17 after 43 generations' insecticide selection compared to 1.00 as sensitive isolate. The nonspecific esterase(NSE) activity of the mosquitoes became strengthened with the extension of the generations, and the individual frequency of those with OD values no less than 0.9 increased gradually, consistent basically to the bioassay. The AChE average inhibition rate decreased with the extended generation and increased resistance, and the individual frequency of those with inhibition rate less than 30% became strengthened with the extension of generations, showing a positive correlation. Conclusion The activity of NSE and AChE shows a correlation with DDVP resistance.

By means of genetic cloning and recombinant techniques, full genome cDNA sequences of rotavirus strain TB-Chen were isolated from an infantile hospitalized with acute gastroenteritis. Nucleotide sequences analyses showed that the full genome of strain TB-Chen contains 18613 nucleotides, encoding 5791 amino acids. Genotyping results showed that the strain TB-Chen belongs to genotype G2P[4]/NSp4[A]. This is the first report on a full genome of Group A rotavirus in China, and has important significance for deep understanding structure and functions of rotaviruses and developing rotavirus vaccines.

In early 2011, the serious outbreak of porcine pseudorabies virus (PRV) infection suddenly recurred in Henan and neighboring Provinces. To investigate the etiology of massive infection with PRV, 16 800 serum samples, 905 porcine epidemic diarrhea virus (PEDV) back-feeding tissues, and 56 PR gene deleted live vaccines were colleted from January 2011 to May 2013 to detect PRV field infection using a PRV gE antibody test kit. The gE and TK genes of 11 new epidemic PRV strains were sequenced by PCR, and their molecular characteristics were analyzed. Moreover, virus titer determination, protective test against PRV, and vaccine potency testing were performed. The results showed that the detection rate of PRV field infection-positive pig farms was 68.06%, and the overall positive rate of PRV field infection in serum was 38.47%; the positive rates in breeding sows, breeding boars, reserve pigs, and commercial pigs were 40.12%, 30.88%, 54.67%, and 26.52%, respectively. The new epidemic strains were in the same evolutionary branch and belonged to the virulent strain group. Compared with the classical PRV strain, the virulence of new epidemic strains changed a little. The length of gE gene was 1 787 bp, and the length of TK gene was 963 bp. The nucleotide homologies of gE and TK genes to Chinese reference strains were 98.2%-99.8% and 98.90%-99.6%, respectively, and the amino acid homologies were 97.1%-99.8% and 97.5%-99.4%, respectively. Commercial vaccine had a 100% protective effect against the new epidemic strains. The positive rate of PRV field infection was 0% in vaccine and 40.44% in back-feeding tissues. The results confirmed that PRV field infection rates were rising sharply among pigs in Henan and neighboring Provinces after 2011. The main virulence genes of new epidemic PRV strains did not change significantly over the years. PR gene deleted live vaccines had no PRV field infection and could completely resist the attack of new strains. The virus carriage of breeding boars and reserve pigs and the serious PRV field infection in PEDV back-feeding tissues were the main causative factors for massive infection with PRV and epidemic outbreak in Henan and neighboring Provinces from 2011 to 2013.
Amino Acid Sequence ; Animal Feed ; analysis ; virology ; Animals ; China ; epidemiology ; Epidemics ; Female ; Herpesvirus 1, Suid ; chemistry ; classification ; genetics ; isolation & purification ; Male ; Molecular Sequence Data ; Phylogeny ; Pseudorabies ; epidemiology ; virology ; Sequence Alignment ; Sequence Homology, Amino Acid ; Sus scrofa ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; chemistry ; genetics

Encephalomyocarditis virus (EMCV) is a natural epidemic zoonotic pathogen. However, no reports have been published regarding the isolation, identification and full-length genome of EMCV from a local aardvark population. In present study, an EMCV isolate HNXX13 was isolated from aardvarks named Huainan-pig in Henan Province. The systematic identification, full-length genome sequencing and molecular characteristic analysis of the isolate HNXX13 were conducted. The result showed that the isolate was spherical with a diameter of 24-30 nm, neither heat- nor acid-resistant, sensitive to trypsin, insensitive to chloroform, not protected by bivalent cationic, and the specific fluorescence was observed in the cytoplasm of BHK-21 cells infected with the isolate by using indirect fluorescence assay. The full-length genome of EMCV HNXX13 generated a 7 725bp sequence (GenBank: F771002), with 81.0%-99.9% nucleotide identity to reference strains from different animals, and 99.5% with a Chinese reference strain isolated earlier from a commercial pig herd. The phylogenetic tree based on the full-length genome and ORF sequences identified that all EMCV strains were divided into three groups G1, G2 and G3, and strain HNXX13 belonging to the G1 group with other Chinese reference strains. The result also identified that this EMCV infection could cause severe clinical signs in a local aardvark population, and enriches the molecular epidemiological data of EMCV in China. Regional differences exist in EMCV genome and transmission is limited within a certain area. However, the cross-infection and transmission of EMCV between aardvark and mice appears most likely. Mutations have occurred in some amino acids of EMCV strain HNXX13 during the transmission in local aardvark herd and these mutations might make the virus easier to infect the aardvark.
Animals ; Animals, Wild ; virology ; Cardiovirus Infections ; veterinary ; virology ; China ; Encephalomyocarditis virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Mice ; Molecular Sequence Data ; Phylogeny ; Xenarthra ; virology