Objective To investigate the mechanisms modulating the functions of dendritic cells (DCs) and suppressing the activation and proliferation of T cells by transforming growth factor-β (TGF-β).Methods Mouse splenic DCs were purified with CD11c+ immunomagnetic beads and the purity of isolated DCs were detected by flow cytometry.Gene chip was used to detect gene expression in DCs after stimulation with TGF-β, and then real-time PCR was performed to analyze the differentially expressed genes in microarray at mRNA level.The activation and proliferation of CD4+ T cells which were co-cultured with DCs after stimulation with TGF-β were detected by flow cytometry.Results The purity of DCs reached over 95% after isolation.TGF-β down-regulated the expression of cell surface markers CD53, CD69, CD33, CD74 and CD93 on DCs;decreased the expression of chemokines Ccl3, Ccl5, Ccl9, Ccl6, Ccl17, Cxcl10, Ccl22, Ccl4, Ccr7, Ccl2, Cxcl9 and Ccl7;inhibited the expression of inflammatory cytokines IL-2ra, IL-12rb2, IL-15ra, IL-1b and IL-15.Moreover, the DCs-mediated activation and proliferation of CD4+ T cells were suppressed by TGF-β.Conclusion TGF-β inhibits the DCs-mediated activation and proliferation of CD4+ T cells by suppressing the expression of surface markers on DCs and down-regulating the expression of chemokines and inflammatory cytokines.

18 fungal strains including N.coenophialum,N.lolii, N.huerfanum、N.chisosum、N.aotearoae、N.sp.and 8 varieties of grass seeds belonging to Festuca arundinacea and Lolium perenne have been studied.With amplification of IS1～IS3 and F1～R1 of genomic DNA, the primers Tub-2-F～Tub-2-R from Tubulin-2 gene and F3～R3 from NC25 gene have been designed.A PCR method to detect N.coenophialum and N.lolii was established, and also a nested-PCR method to detect N.coenophialum and N.lolii in single seed was established.These PCR detection methods are strongly special and much credible and rapid-speeded.

Background Researches determined that the alteration of A69S locus of age-related maculopathy susceptibility 2 ( ARMS2 ) gene is closely associated with the pathogenesis and progression of age-related maculopathy ( AM D ).However,the location of ARMS2 protein in normal eye tissue is still in controversy,therefore,its function is below understanding up to now.Objective The goal of this laboratory work was to investigate the distribution,expression and location of ARMS2 protein in normal adult retina and choroid as well as in retinal pigment epithelial (RPE) cells and lay a basis for exploring further its function in the protein level.Methods Ten donor eyeballs of normal adult male with the age from 28-42 years were collected in eye bank of Qingdao Eye Hospital.The frozen sections of the retina and choroid were prepared for the detection and location of ARMS2 in 3 eyes by immunofluorescence under the confocal laser microscope.The retina was isolated for the primary culture of RPE cells using explant culture method.The cells were then identified by CK32 antibody by immunofluorescence.The distribution and expression of the ARMS2 protein in retina,ehoroid and RPE cells were determined by immunofluorescence technique.Results ARMS2 protein was strongly expressed in retinal vessel,RPE cell layer,Bruch membrane and choroidal vessel,but weak expression was in retinal ganglion cell layer,inner nuclear layer,outer plexiform layer,outer nuclear layer and inner plexiform layer in the normal eyes.The primarily cultured cells appeared the polygon shape with the abundant pigment in cytoplasm.The immunofluorescence of the cells showed the positive response for CK32,exhibiting the green fluorescence granules in the cytoplasm.The positive expression of ARMS2 protein also was seen in the cytoplasm of RPE cells,appearing the red fluorescence.Conclusions ARMS2 protein mainly distribute and locate retinal and choroidal vessels,RPE cells and Bruch membrane in normal eye.

Objective To observe the clinical efficacy of butylphthalide soft capsules for treatment of acute cerebral infarction and its effect on the neurological function and inflammatory reaction of patients.Methods A total of 100 patients with acute cerebral infarction in the First Hospital of Zhangjiakou City and the Third Hospital Affiliated to Hebei North University from January 2016 to October 2016 were selected and divided into control group (n =50) and treatment group (n =50) according to the treatment protocols.The patients in the control group were given anti-platelet aggregation,statins,and other routine treatment;based on this,the patients in treatment group were orally administrated with butylphthalide soft capsules.The elbow venous blood of patients in the two groups was collected and the C reactive protein(CRP),interleukin-6 (IL-6) levels,the platelet aggregation rate were detected.The change of National Institute of Health stroke scale(NIHSS) score of patients after four weeks treatment was compared between the two groups.The recovery of neurological function of patients in the two groups was evaluated by modified Rankin grading after three months treatment.Results There was no statistic difference in the CRP,IL-6 levels and platelet aggregation rate before treatment between the two groups (P ＞ 0.05).The CRP,IL-6 levels and platelet aggregation rate of patients in the two groups after one week treatment were significantly lower than those before treatment (P ＜ 0.05);and the CRP,IL-6 levels and platelet aggregation rate of patients in the treatment group were significantly lower than those in the control group after one week treatment (P ＜ 0.05).The NIHSS score of patients in the treatment group was significantly lower than that in the control group after four weeks treatment (P ＜ 0.05);the Rankin score of patients in the treatment group was significantly lower than that in the control group after three months treatment (P ＜ 0.05).The total effective rate of patients in the treatment group was significantly higher than that in the control group (x2 =13.22,P ＜ 0.05).Conclusion Butylphthalide soft capsules can significantly reduce the inflammatory reaction in patients with acute cerebral infarction,improve and recover the neurologic impairment;and its curative effect is remarkable.

Objective This study was performed to investigate the levels of HSP70 in tissue in pemphigus as a possible new theoretical basis for further elucidate the pathogenesis of pemphigus.Methods The expression of HSP70 in 62 patients with pemphigus was determined by immunohistochemistry,and the normal skin was taken as control.Results The results showed that the positive cells of HSP70＞75 ％ in the blisters of pemphigus vulgaris and the positive cells of HSP70＞50％ in the inflammatory cells near the blisters,and the expression of HSP70 was significantly higher than that in normal skin,which was statistically significant(Z=5.42,4.73,P＜0.01).Conclusion The abnormal expression of HSP70 in inflammatory cells and psoriasis of pemphigus patients showed that HSP70 is involved in the pemphigus.

OBJECTIVETo study the effect of dimethoate on the expression of heat shock protein 70 (HSP70) in peripheral blood lymphocytes of human beings and to explore the feasibility of HSP70 in biomonitoring among workers exposed to organophosphorous pesticides.

METHODSPeripheral blood lymphocytes were isolated from subjects, comprising 11 people of the control group and 35 workers of the exposure group exposed to dimethoate. Flow cytometry was used for detecting both the basic level and the level of the dimethoate-induced expression of HSP70. The activity of whole blood acetylcholinesterase (AChE) was examined at the same time. Then the potential influential factors to HSP70 expression and AChE activity were analyzed.

RESULTSThe basic level of HSP70 expression in the exposure group and the control group was 41.24% +/- 10.45% and 23.97% +/- 4.29% respectively. The activity of AChE in these two groups were (125.23 +/- 7.97) and (145.36 +/- 8.78) U/ml respectively. Both differences were statistically significant (P < 0.01). Among the exposure group, the basic level of HSP70 expression of the two categories comprising operators and packers, were 47.34% +/- 11.87% and 38.05% +/- 8.20% respectively (P < 0.05), while there was no significant difference (P > 0.05) in AChE activity between these two categories. The factors that had significant influence on the HSP70 basic level of the exposure group were the health condition, the environmental concentration of dimethoate and the exposure time in order, according to their significance of influence. At least 88% variance of HSP70 could be explained by these factors. The only factor that could influence AChE activity significantly was the exposure time, and it could only explain about 12% variance of AChE activity. After the treatment of dimethoate in vitro, the level of the induced expression of HSP70 in the control group was significantly higher than that of the exposure group (P < 0.01). The increasing order was the control group, the group of packers and the group of operators according to the increasing extent and there were significant difference among them (P < 0.01). The factors that could significantly influence the change ratio of HSP70 expression were the environmental concentration and the exposure time.

CONCLUSIONHSP70 is a potential index that can reflect the individual and environmental conditions of workers exposed to dimethoate comprehensively.

Acetylcholinesterase ; blood ; Adult ; Cells, Cultured ; Dimethoate ; toxicity ; Female ; HSP70 Heat-Shock Proteins ; biosynthesis ; Humans ; Insecticides ; toxicity ; Lymphocytes ; drug effects ; metabolism ; Male ; Middle Aged ; Occupational Exposure

0.05).However,in the end of the treatment,the patients in the treatment group scored significantly better with all the scales than the control group(P

The influenza A virus matrix protein2 gene(M2)which deleted transmembrane region was amplified by overlap extending PCR,and the multiepitope gene of hemagglutinin(HA)was PCR amplified with seven continuous synthesized segments by designing primer.The two gene segments were separately cloned into pMD18T vector to sequence analysis and prokarytic expression vector pET28a+ to construct the recombinant plasmid pET28a+M2dHA.The recombinant plasmid was transformed into E.coli BL21(DE3),and the high expression strain was obtained by screening monoclones.The recombinant protein existed as inclusion bodies,which accounted about 45% of the total cellular protein.The inclusion bodies were washed with 1% Triton X100 solution twice,and dissolved in 8 mol/L urea solution.The solution protein was purified by Ni+2 affinity chromatography,and refolded by dilution renaturation,then purified by Q Sepharose FF cation exchange column.The purity of the protein was over 90% by HPLC analysis.The result of Western blot showed it has good antigenicity and specificity.These results strongly supported for the further study of the broadspectrum influenza virus subunit vaccine.

Background Endophthalmitis is a serious,sight-threatening condition.Identifying the causative microorganisms is very important for available treatment of endophthalmitis.Objective This survey was to analyze the spectrum of organisms causing culture-proven endophthalmitis and their sensitivities to commonly antimicrobial agents.Methods Medical data of patients with culture-proven endophthalmitis at Zhongshan Ophthalmic Center from January 2003 through December 2010 were respectively reviewed.The outcomes included intravitreal isolates and antibiotic sensitivities were analyzed.Results Four hundred and sixty-nine strains of organisms were isolated from 447 eyes of 447 patients with infective endophthalmitis,including 22 eyes of polymicrobial infection.In the organisms,gram-positive organisms were 241 (51.4％),fungi were 125 (26.7％) and gram-negative organisms were 103 (22.0％).The most common organisms were Staphylococcus epidermidis in 29.4％,Aspergillus in 7.7％ and Pseudomonas aeruginosa in 5.3％.In this group of infective patients,the most common clinic settings were posttraumatic endophthalmitis (72.7％),and then were postoperative endophthalmitis (10.5％),endogenous endophthalmitis (9.8％) and keratitis (6.9％).Most gram-positive organism and gram-negative organism were sensitive to levofloxacin and cefoperazone.The susceptibility rate of gram-positive organism to chloromycetin was increased in 2007-2010 years compared with 2003-2006 years (x2=5.398,P＜0.05).The susceptibility rate to ciprofloxacin of gram-negative organisms declined (x2 =5.398,P ＜ 0.05),but that to rifampicin increased in the duration of 2007-2010 compared with 2003-2006 (x2 =4.500,P ＜ 0.05).Conclusions Gram-positive organisms are the most commonly causative organisms of endophthalmitis.Most bacterial organisms are sensitive to levofloxacin and cefoperazone.Local data of culturing and susceptibility test offers a guideline for the treatment of infectious endophthalmitis.

OBJECTIVETo explore the value of fluorescence in situ hybridization (FISH) technique in diagnosis of variant Ph chromosome translocation (VT) and Ph chromosome-negative chronic myelocytic leukemia (CML).

METHODSNine CML patients with VT and 2 Ph chromosome-negative CML patients confirmed by R banding were assayed with dual color-dual fusion BCR/ABL probe by FISH.

RESULTSThe 9 patients with VT involved chromosomes 1, 3, 5, 12, 13, 15, 17 and 21 besides chromosomes 9 and 22, and some of them showed recurrent aberrations; FISH results were positive and the signal feature was 2R2G1Y. The 2 Ph-negative CML patients had normal karyotypes; FISH was positive and the signal feature was 1R1G2Y and 1R1G1Y respectively.

CONCLUSIONFISH can provide better diagnosis for CML with VT and Ph-negative CML. Abnormal karyotype and marker gene changes can be assessed based on the signal feature of the positive cell. So FISH is a complementary method to banding technique in diagnosis of CML.

Adult ; Aged ; Chromosomes ; genetics ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; genetics ; Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative ; diagnosis ; genetics ; Male ; Middle Aged ; Philadelphia Chromosome ; Translocation, Genetic ; Young Adult