Emergencies ; Food Services ; organization & administration ; Foodborne Diseases ; prevention & control ; Humans ; Risk Assessment

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; Cells, Cultured ; Female ; Gene Expression ; drug effects ; Neurons ; drug effects ; Phenylalanine ; toxicity ; Phenylketonurias ; complications ; Rats ; Rats, Sprague-Dawley ; rho GTP-Binding Proteins ; genetics

Objective:To investigate the effects of shikonin on osteoclastogenesis in vitro and amelioration of bone loss in ovariectomized-induced osteoporosis in mouse model.Methods:The optimal concentration of shikonin treating were evaluated in vitro depending on its effect on the viability of C57BL/6J mouse bone-marrow-derived macrophages by CCK-8 method.To establish the osteoclastogenesis cell model,macrophages were cultured with RANKL and M-CSF treatment,and TRAP staining was used to observe the generation of osteoclasts after treating with different concentration of shikonin solution.Expressions of osteoclast marker genes,including TRAP,c-Fos and NFATclwere detected with real-time PCR.Fifthteen mice were randomly allocated into sham operation group,ovariectomized model group and shikonin treatment group.After the modeling,mice in treatment group were received the intraperitoneal injection of shikonin,while the other two groups treated with normal saline.After thirty days treatments,all animals' tibias were dissected for micro-CT analysis.Results:①The macrophages viability was significantly inhibited when the concentration of shikonin was higher than 250 nmol/(P＜0.01).②The osteoclastogenesis was significantly suppressed by differemt dose of shikonin(P＜0.01).③ The expression of the osteoclastic marker genes (TRAP,c-Fos and NFATc 1) were suppressed by addition of shikonin comparing to control group (P＜0.01).④ Shikonin effectively prevented ovariectomy-induced bone loss (P＜0.05).Conclusion:Shikonin suppresses osteoclastogenesis in vitro and ameliorates ovariectomized-induced osteoporosis in mouse model.

Objective To study the clinical significance of the expression of D-amino acid oxidase(DAO) in hepatocellular carcinoma(HCC) tissues.Methods The gene expression profiles and related clinical data of 214 HCC cases were collected.Univariate and multivariate analyses were conducted to investigate the association between DAO expression levels,clinical traits,the prognosis.Results Univariate analysis indicated that there were a lower level of blood alpha fetoprotein(AFP,P=0.001),a smaller number of nodules(P=0.042),a better TNM stage(P=0.014),a lower metastasis risk(P=0.001) and a better prognosis(P=0.011)in the samples with high DAO expression.Multivariate analysis also indicated a lower AFP level(OR:0.162,95%CI:0.078-0.336) and metastasis risk(OR:0.140,95%CI:0.069-0.284) in the samples with high DAO expression,as well as a better prognosis(OR:0.833,95%CI:0.700-0.992).Conclusion DAO expression was associated with blood AFP levels,metastasis risk and prognosis in HCC,and its high expression was a protective factor for HCC.

Objective To clone and express translocation intimin receptor(Tir)of enterohemorrhagic Escherichia coli(EHEC)O157:H7,and to analyze the effect of different routes on the induction of immunity to the recombinant protein.Methods The tir eucoding genes were amplified from EHEC O157:H7 strain guangzhou 246 genome,and genes were cloned into the vector pET-30a(+).The pET-30a(+)tir recombinant was transformed into E.coli BL21.and expression was induced bv IPTG.The expressed product was analyzed by SDS-PAGE and purified by Ni-IDA affinity chromatography.The immunized mice sera and fecal against the recombinant protein was detected.Resuits The length of the tir is 1677 bp,with the initiation codon ATG and the termination codon TAA.Double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid pET-30a(+)-tir was constructed.The recombinant protein was expressed in Escherichia coli expression system,and was purified by Ni-IDA affinity chromatography.The mice were able to produce a high serum IgG antibody titer after both subeutaneous and intranasal immunizations.Meanwhile,the intranasal immunization induced serum and fecal IgA antibody titer was significantly higher than that of the subcutaneous immunization group.Conclusion Tir molecule is potential vaccine candidate for preventing EHEC disease.

Objective To analysis the clinical manifestations of a large Spinocerebellar Ataxia 3 pedigree to pro-vide the information for the early diagnosis of Ataxia 3. Methods SCA3/ATXN3 gene was determined by using Poly-merase Chain Reaction and fragment analysis in the large pedigree members and patients ’clinical data was collected. Five patients underwent MRI imaging and fundus examination. Results There were eighteen clinical patients and twelve ATXN3 carriers in this Pedigree . In addition to ataxia, three patients presented with intellectual disability, one with cer-vical spondylosis, one with dysmyotonia, one with disorder in visual system, and seven with abnormality in autonomic ner-vous system. The MRI revealed that pons and cerebellar atrophy in some patients inordinately. Undus examination did not reveal any obvious abnormality. Conclusions The symptoms of SCA3 are heterogeneous in the same pedigree. When patients present with symptoms of cerebellar system, visual system and autonomic nervous system, or cervical spondylosis and intellectual disability, SCA3 should be considered.

Objective To analyze the clinical effect of biliary-enteric anastomosis and biliary stent to palliative treatment of malignant obstructive jaundice.Methods A total of 40 patients with inoperable malignant obstructive jaundice were enrolled in this study,and 20 patients were performed biliary stent placement (stent group),simultaneously 20 patients were performed biliary-enteric anastomosis (operation group).The fatality rate after operation,the level of total bilirubin before treatment and after treatment for 4,7,14 d,the rate of hyperpyrexia,nausea and vomiting,postoperative recurrence of jaundice were compared between two groups.Results There were no dead in two groups.The level of total bilirubin was decreased after treatment,and there was no significant in two groups before treatment and after treatment for 4,7,14 d (P ＞ 0.05).The rate of hyperpyrexia in operation group was significantly lower than that in stent group (0 vs.4/20) (P ＜ 0.05).Conclusion For palliative treatment of malignant obstructive jaundice,the biliary-enteric anastomosis should be performed first if there is no significant contraindication.

OBJECTIVETo explore efficacy enhancing and detoxification roles of Jiedu Quyu Zishen Recipe (JQZR) in treating systemic lupus erythematosus (SLE) by studying its effect on Toll like receptor 9 (TLR9) signal pathway of murine macrophage cells after JQZR stimulated CpG oligodeoxynucletide (CpG ODN).

METHODSMurine macrophage cells in vitro cultured were randomly divided into 4 groups, i.e., the blank serum group, the CpG ODN stimulus group, the CpG ODN + dexamethasone group, the CpG ODN + medicated serum group. Murine macrophage cells were collected after 24-h intervention. The expression of TLR9, myeloid differentiation factor 88 (MyD88), NF-KB, IFN-α mRNA were analyzed by RT-PCR. The expression of TLR9 and NF-κB protein were analyzed by Western blot. Changes of the NF-KB transcriptional activity were assayed by Dual-Luciferase reporter assay system.

RESULTSmRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were enhanced, showing statistical difference when compared with those of the blank serum group (P <0. 05, P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of MyD88, NF-κB, and IFN-α, the protein expression of NF-κB and the NF-κB transcriptional activities decreased in the CpG ODN + dexamethasone group with statistical difference (P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were decreased in CpG ODN+ medicated serum group with statistical difference (P <0. 01).

CONCLUSIONEfficacy enhancing and detoxification roles of JQZR in treatment of SLE might be realized through regulating TLR9 signal pathways.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Macrophages ; metabolism ; Mice ; Myeloid Differentiation Factor 88 ; NF-kappa B ; RNA, Messenger ; Signal Transduction ; Toll-Like Receptor 9 ; metabolism

Four strains of aerobic denitrifying bacteria were isolated from biomembrane of the laboratory biofilter by selective medium. The denitrifying rates of these four strains were found more than 80% under aerobic condition for 40 hours. The denitrifying rate of A1 was highest, which was 99.05%. When measuring the course of nitrogen element changing, it is found that they accumulated nitrite expect the strain A1. The strain A1 was gram positive and spherical. It is identified as Pseudomonas putida based on its biochemical and morphological characters and phylogenetic analysis of 16S rDNA sequences(genbank accession NO.DQ836052.1). For the strain A1, the optimum beginning pH was 7.0 around, and the optimum tempera- ture was 30 ℃ around, The change of DO did not influence the effect of denitrification when it was more than 2.0 mg/L.

Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome.The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique.The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer.The extraction was tested for four activated sludge differing in places and dates.The DNA fragment from all sludge was integrity.DNA yields ranged from 105 to 823 ?g/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested.In general,all methods produced DNA pure were not enough for PCR amplification and libraries construction.As basis of experimental goals,the study provides an appropriate extraction method of microbial DNA in sludge.