We investigated the carrying situation of Escherichia albertii from healthy people engaged in breeding and slaughtering poultry for a long time.We collected stool samples from people engaged in breeding and slaughtering poultry and other healthy people.After enriched with EC broth,eae-positive enrichment culture was directly streaked on MacConkey,and eaepositive lactose non-fermenting isolate was retained for further investigation.The 16S rDNA sequencing and multilocus sequence typing (MLST) were applied in the identification of E.albertii from suspected strains.Intimin subtypes and cdtB types of E.albertii strains were detected.Pulsed-field gel electrophoresis (PFGE) was used to detected genetic polymorphism of strains from this study and animal source ones.Results showed that two isolates were identified as E.albertii from 189 stools of people exposed to slaughtering chickens and ducks and one from 58 stools in control groups.No isolate was identified as E.albertii from 138 stools samples of people exposed to breeding poultry.Intimin subtypes of three isolates from stool samples were subtyped as sigma,iota 2,nu,and cdtB types were closely related to types Ⅱ/Ⅲ/Ⅴ.PFGE patterns of the three strains was distinguishable (＜80％ similarity),and appeared in different cluster with chickens,ducks and other sources of E.albertii strains.The rate of carrying E.albertii to a certain extent exist in healthy people engaged in slaughtering chickens and ducks,and the relationship between these strains and strains from poultry should be further investigated.

Objective To explore the effect of blocking BTLA-HVEM (herpesvirus entry mediator-B and T lymphocyte attenuator) pathway on dendritic cell function and the related immunological mechanisms. Methods Murine BTLA extracellular domain eukaryotic expression vector psBTLA was constructed by gene recombination and transfected CHO by Lipofection method. Mouse bone marrow cells were induced to differentiate into DCs by GM-CSF plus IL-4. Expression of BTLA and HVEM on DCs was detected after HSPT0-TC-1 peptide complex stimulation by FACS. Expression of BT-1 and secretion of IL-12 were detected after HSP70-TC-1 peptide complex plus psBTLA transfected CHO culture supernatant stimulation on DCs. Pretreated DCs co-cultured with the same genetic background mouse splenocytes and lymphocytes proliferation and cytokine secretion were detected. Effect of psBTLA gene transfer in vivo on BT-1 expression of DCs and tumor growth on tumor-bearing mice was detected. Results Extracellular domain of murine BTLA was successfully constructed, psBTLA stable transfection CHO cells were obtained and expression of BTLA extracellular domain(sBTLA) was detected the in its culture supernatant. BTLA and HVEM expression of DCs were increased after stimulation by the antigen peptide complex. When DCs were treated with antigen peptide complex plus culture supernatant containing sBTLA, B7-1 expression and IL-12 secretion were increased. Co-cultured with splenocytes, lymphocytes proliferation and cytokine secretion, such as IL-2 and IFN-γ,, were also increased. Gene transfection with psBTLA in vivo promoted B7-1 expression on DCs and inhibited cervical cancer cells growth. Conclusion Blockade of BTLA-HVEM inhibitory pathway with sBTLA can further improve DCs function, activation of lymphocytes and promote antitumor immune response.

Objective To study the effects of selenium deficiency,iodine deficiency and combined selenium and iodine deficiency on bone and cartilage growth in the parental and the first filial generation rats. Methods Forty-eight weanling healthy SD rats were randomly divided into selenium deficieney, iodine deficiency, combined selenium and iodine deficiency and control groups according to their body mass. These rats were fed with selenium deficiency, iodine deficiency, combined selenium and iodine deficiency, and normal fodder, respectively. The parental rats (about 3 months old) were mated in each group 8 weeks after the beginning of the experiment. Right tibias and left knee joints were collected when the parental generation rats were about 6 months and the first filial generation rats were about 3 months old. Tibial length, mid-shaft tibial diameter, and articular cartilage diameters of the right tibias were measured by vernier caliper. Left knee joints were embedded and cut into sections and the thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in growth plate cartilage were observed under the light microscope. Results The selenium deficiency had significant effect on serum selenium level of the parental and the first filial generation rats(F value were 239.56,232.68, P< 0.01), and also on serum T4 level of the first filial generation rats(F value were 6.95, P < 0.05). The iodine deficiency had significant effect on serum T3 and T4 level in the two generations rats(F value were 14.11,14.05,30.29,34.53, P < 0.01 ). There were interactions between selenium deficiency and iodine deficiency on serum T4 level in the first filial generation rats (F= 5.99, P< 0.05). The serum selenium of selenium deficiency group[ (30.28 ± 6.34), (43.95 ± 9.75)μg/L],combined selenium and iodine deficiency group[ (30.33 ± 5.18), (35.40 ± 3.16)μg/L] were significantly lower than iodine deficiency group[(345.83 ± 29.55), (245.24 ± 9.95)μg/L] and the controls[ (358.64 ± 30.50), (236.50 ±9.75) μg/L] in the two generations. The serum T3 of combined selenium and iodine deficiency group [(0.55 ± 0.05 ),(0.88 ± 0.14)nmol/L] were significantly lower than the controls[(0.75 ± 0.08), (1.26 ± 0.26)nmol/L] in the two generations. The serum T4 of iodine deficiency [ (24.11 ± 2.29), (42.10 ± 8.92) nmol/L ] and combined selenium and iodine deficiency group[ (20.66 ± 1.93), (26.55 ± 5.98)nmol/L] were significantly lower than the controls[ (36.15 ±2.74), (52.79 ± 8.84)nmol/L] and selenium deficiency group[ (28.12 ± 3.33), (52.02 ± ll.99)nmol/L] in the two generations. The selenium deficiency and iodine deficiency had significant effect on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in first filial generation rats(F values were 24.31,6.98,40.76,56.15,25.24,82.82, 10.07,5.57, P <0.05 or <0.01). There were interactions between selenium deficiency and iodine deficiency on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes (F values were 5.68,24.86,41.82,9.12, P <0.05 or <0.01 ). The tibial length of the selenium deficiency group[ (33.17 ± 0.34)mm] and combined selenium and iodine deficiency group[ (31.30 ± 0.87)mm] were significantly lower than the controls[ (34.12 ± 0.32)mm, P< 0.05]. Thickness of the growth plate cartilage [ (1.60 ± 0.18)mm ], layers of proliferative chondrocyte (8.54 ± 0.81), and hypertrophic chondrocyte (4.95 ± 0.37)of the combined selenium and iodine deficiency group were significantly decreased when compared to the selenium deficiency group[ (3.03 ± 0.10)mm, 14.68 ± 0.84,6.60 ± 0.31], iodine deficiency group[ (2.90 ± 0.09)mm, 13.75 ±0.33,6.61 ± 0.84 ] and the controls [ (3.19 ± 0.09) mm, 14.94 ± 0.36, 6.64 ± 0.26, P <0.05]. Thickness of the growth plate cartilage, layers of proliferative chondrocyte of the iodine deficiency group were lower than the controls(P<0.05). Conclusions Selenium deficiency impair tibial growth in first filial generation rats, iodine deficiency retarded the chondroncyte proliferation and decreases the thickness of growth plate cartilage in first filial generation rats, and combined selenium and iodine deficiency significantly impair the growth of bone and cartilage in first filial generation rats.

Objective To master the level of iodine nutrition among population in Jiangxi province,and to provide a scientific basis for establishing the strategy for prevention and control of iodine deficiency disorders (IDD).Methods Retrospective method was adopted to analyze the goiter rate and frequency distribution of urinary iodine of children aged 8-10,the qualified rate of iodized salt,the coverage rate of iodized salt and the consumption rate of qualified iodized salt in residents of Jiangxi province from 1995 to 2010.The method of correlation analysis was used to analyze the relationship between goiter rate of children (by palpation) and the qualified rate of iodized salt,iodized salt coverage rate and residents consumption rate of qualified iodized salt.Results The goiter rates (measured by the method of palpation) of children aged 8-10 were down from 40.17％(482/1200) in 1995 to 0.80％(16/2000) in 2010(x2 =4.864,P＜ 0.05).The median of urinary iodine of children was higher than 200 μg/L; the proportion of people whose urinary iodine content higher than 300 μg/L was above 25.00％ and the highest propoaion was up to 58.01％ (210/362) between 1995-2010.The minimum median of salt iodine was 17.77 mg/kg in 1995,and 29.30-39.10 mg/kg in other years.The qualified rates of iodized salt,the iodized salt coverage rates and the consumption rates of qualified iodized salt increased from 43.58％(452/1037),86.42％(1037/1200) and 37.67％(452/1200) in 1995 to 97.95％ (1916/1956),99.95％(1956/1957) and 97.90％(1916/1957) in 2010,respectively; there was a growth trend over the years(x2 =5.240,6.118,5.631,all P ＜ 0.05).The goiter rates of children were related to the qualified rates of iodized salt,the iodized salt coverage rates and the consumption rates of qualified iodized salt,and the correlation coefficient(r) was-0.833,-0.881 and-0.918 (all P ＜ 0.05),respectively.Conclusions The level of iodine nutrition among residents in Jiangxi province has already gone beyond the appropriate level,and the iodine concentration in salt should be cut to ensure the appropriate iodine nutrition level among people.

Objective To investigate the influence of Zhenqing Recipe(ZQR)and Ligustri Lucidi Fructus(LLF)on diabetic rats and its possible mechanism.Methods The model of type 2 diabetic rats was established by feeding a high-sucrose-high-fat diet and injecting a low dose of Streptozotocin in Wistar rats.The model rats were randomly divided into three groups: diabetic model,ZQR-treated,and LLF-treated groups for 8-weeks treatment.The normal Wistar rats were as a normal control group.Results The level of fasting blood glucose in ZQR and LLF groups was decreased compared with model group(P < 0.01,0.05,respectively).Both ZQR and LLF markedly reduced serum triglycerides(P < 0.01,0.05,respectively),and increased the insulin sensitivity index(P < 0.05).Histopathology revealed that ZQR and LLF reduced pancreatic damage.Immunohistochemistry evaluation showed that the percentage of insulin positive cells in pancreatic island was higher than model group(P < 0.01,0.05,respectively).The mRNA and protein expression of SREBP-1c in pancreas were significantly decreased in ZQR and FLL group(P < 0.01).Conclusion ZQR has therapeutic effect on type 2 diabetes,it ameliorates the histopathologlcal changes of pancreas,protects β cells,improves insulin resistance,and attenuates the expression of SREBP-1c.This study also provides the anti-diabetic evidence of FLL even its effects are weaker than ZQR.

Objective To evaluate five years results of laser in situ keratomileusis(LASIK)for correction of myopia.Design Ret- rospective case series.Participant 64 patients(126 eyes)with myopia received operation of LASIK.Method 126 myopic eyes were treated with LASIK.Based on the preoperative spherical equivalent refraction(SER),the patients were divided into Group A,B and C(-3.00D～6.00D,-6.25D～-10.00D and -10.38D～-19.88D).Before and after operation,the visual acuity,refraction,anterior seg- ment,funduscopy,intraocular pressures of patients were measured,and the patients were followed-up five years.Main Outcome Measure Visual acuity,intraocular pressure,refraction and complication.Result At five years,in Group A,B,C,uncorrected visual acuity was≥1.0 in 87.2%,69.1%,31.3% patients,and 100%,98.2%,75% had vision of≥0.5.After five years of operation,the re- fraction was -0.70D?0.52D,-1.06?0.13D,-2.46?2.14D in three groups respectively,there was no significant difference between Group A and B(P=0.23);But there was significant statistical difference between Gruop A and C(P=0.008),Group B and C(P=0.021)respec- tively.At the fifth-year,68.3%,45.0% and 23.5% of refraction in Group A,B,C were within?1.00D.Intraocular pressure at the fifth year was 11.91?2.35mmHg,11.31?2.20 mmHg and 9.24?2.20 mmHg respectively.In Gruop B and C,one patient complained glare re- spectively.In Gruop B,one patient's intraecular pressure was elevated after using glueocorticoid.In Group C,one eye was complicated with rbegmatogenous retinal detachment at the second year after LASIK.Conclusion In the follow-up of five years,LASIK is effective and safe.The results in middle and high myopia are better than that in extreme high myopia.(Ophthalmol CHN,2006,15:312-314)

Animals ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Endothelial Cells ; cytology ; Hepatocytes ; cytology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the genetic pathogenesis of myasthenia gravis (MG) caused by cytotoxic T lymphocyte associated antigen-4 (CTLA-4) gene polymorphisms and regulation function of transcription factor.

METHODSELISA assay was used to determine the expression level of serum sCTLA-4 in MG. Four single nucleotide polymorphisms (SNPs) of CTLA-4 at exon 1 +49, promoter -318, -1661, -1772 were analyzed by restriction fragment length polymorphism (RFLP). Transcription factor nuclear factor 1(NF-1) and c/EBPbeta binding site were confirmed by chromatin immunoprecipitation(CHIP) assay.

RESULTSIt was found that the frequencies of the GG+49 genotype and G+49 allele are higher in MG patients with thymoma than those in patients of thymic hyperplasia and normal thymus subgroups. T/C-318 is not correlated with MG. The frequency of CT-1772 genotype is significantly higher in MG patients, especially in MG patients with thymoma, when compared with that in healthy controls. Meanwhile, the frequency of the G-1661 allele and GG-1661 genotype is lower in MG patients. Linkage disequilibrium (LD) between each SNPs in promoter -1772, -1661, -318 and coding sequence 1 (CDS 1) +49 is apparent. sCTLA-4 levels in patients' sera are correlated with the haplotype and genotype. T/C-1772 and A/G-1661 SNPs change the sequence of transcription factor NF-1 and c/EBPbeta binding sites. DNA variants lose site-specific binding activity of transcription factor regulated by lectin ConA and PHA.

CONCLUSIONThere are strong positive linkages among four SNPs. C/T-1772 and A/G-1661 polymorphisms can result in inefficient transcription of CTLA-4 gene. T>C-1772 mutation also affects gene splicing. These SNPs may constitute a factor of susceptibility to disease.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; Antigens, Differentiation ; blood ; genetics ; CCAAT-Enhancer-Binding Protein-beta ; genetics ; CCAAT-Enhancer-Binding Proteins ; genetics ; CTLA-4 Antigen ; Exons ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; genetics ; immunology ; NFI Transcription Factors ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Thymoma ; genetics ; Thymus Hyperplasia ; genetics ; Thymus Neoplasms ; genetics ; Transcription Factors ; genetics

OBJECTIVETo investigate whether Notch signaling is activated in hepatic stellate cells (HSCs), and to determine whether manipulation of the Notch signaling pathway can effect the activation of HSCs.

METHODSThe expression of Notch signaling components in unactivated or TGF-b1-activated HSC-T6 cells was detected by Taqman Probe-based gene expression analysis. Differential expression of Notch3 and Jagged1 was detected by immunofluorescence analysis. Notch3-mediated expression of the myofibroblastic markers, a-SMA and collagen I, was detected in HSC-T6 cells transfected with pcDNA3.1-N3ICD or Notch3 siRNA by Western blotting.

RESULTSNotch signaling components were expressed in both unactivated and activated HSC-T6 cells, but the TGF-b1-treated cells showed significantly higher expression levels of Jagged1 (3.9-fold, F = 2543.482), Notch3 (4.2-fold, F = 287.982), and HES1 (3.2-fold, F = 1719.851). Transfection-mediated over-expression of Notch3 led to significantly increased expression of a-SMA (6.8-fold, t = 13.157) and collagen I (5.5-fold, t = 9.810) (both P less than 0.01). Transient knock-down of Notch3 expression by siRNA decreased expression of the myofibroblastic markers (a-SMA by approximately 90%, t = 19.863 and collagen I by 84%, t = 10.376; both, P less than 0.01). Moreover, knock-down of Notch3 antagonized the TGF-b1-induced expression of a-SMA and collagen I.

CONCLUSIONNotch signaling may participate in liver fibrogenesis by regulating HSC activation. Selective interruption of Notch3 may represent a new anti-fibrotic strategy to treat liver fibrosis.

Animals ; Calcium-Binding Proteins ; genetics ; metabolism ; Cell Line ; Hepatic Stellate Cells ; metabolism ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Jagged-1 Protein ; Membrane Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Rats ; Receptor, Notch3 ; Receptors, Notch ; genetics ; metabolism ; Serrate-Jagged Proteins ; Signal Transduction

OBJECTIVETo study the effect of butenolide (BUT) on cultured chondrocytes differentiation and the possible protective effects of selenium (Se).

METHODSEx-vivo cultured chondrocytes were divided into six groups: (1) Control group (without BUT and Se); (2) Se 0.1 microg/ml control group; (3) BUT 0.1 microg/ml group; (4) BUT 1.0 microg/ml group; (5) BUT 5.0 microg/ml group; and (6) BUT 1.0 microg/ml + Se 0.1 microg/ml group. The expression of collagen II (Col II), collagen X (ColX), basic fibroblast growth factor (bFGF), and parathyroid hormone-related peptide (PTHrP) in (or around) chondrocytes in all groups were analyzed by immunohistochemistry.

RESULTSThe expressions of Col II in 1.0 microg/ml BUT group and 5.0 microg/ml BUT group were significantly lower than those in the control group (P < 0.05). The expression of Col II in 1.0 microg/ml BUT + Se group were significantly higher than those in the 1.0 microg/ml BUT group and 5.0 microg/ml BUT group (P < 0.05). The expressions of bFGF and PTHrP of BUT groups were significantly higher than those in the Se and control groups (P < 0.05). No expression of ColX was observed in all groups.

CONCLUSIONBUT can affect the collagen II synthesis of the chondrocytes. Selenium supplementation may play a protective role.

4-Butyrolactone ; analogs & derivatives ; pharmacology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Humans ; Protective Agents ; pharmacology ; Selenium ; pharmacology ; T-2 Toxin ; toxicity