After the mice had been injected im203HgCl2 185 kBq/mouse, they were injected ip with chelating agents. There were 15 chelating agents of 3 types,of which the effect of eliminate mercury of pyridoxine derivative, benzenesul-fonate cysteine and sodium dimercaptosuccinate ( DMS-Na) were better than the others. The urine excretion of 208Hg was increased and the ratio were 4.6,11.5 and 22 respectively in comparison with the control group.The minimum 203Hg content in organs of mice was in the DMS-Na group,the next was the benzenesulfonate cysteine group,but there no significant difference between these 2groups.

OBJECTIVE To investigate genes associated with aminoglycosides modification enzymes(AMEs) in Pseudomonas aeruginosa(PAE) isolated from ICU patients.METHODS Drug-reisistant genes encoding AMEs such as aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ were detected by polymerase chain reaction(PCR)(amplification) in 21 PAE isolates.RESULTS The positive rates of aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aac(3)-Ⅰ genes were positive in 19.0%,23.8%,9.5%,4.8%,19.0% and 0% of 21 isolates,respectively. Drug-resistant genes encoding AMEs were detected positively in 42.8% of 21 isolates.(CONCLUSIONS) AMEs genes are present in high percentage of PAE isolated from ICU patients.

Objective To investigate the roles of a variety of cytokines including transforming growth factor beta (TGF-β),interleukin-6 (IL-6) and interleukin-2 (IL-2) in the differentiation of CD+4 Tlymphocyte cells.Methods T lymphocyte cells either in human peripheral blood or routine spleen were cultured in vitro under different stimulation conditions.Flow cytometry was used to analyze the percentages of CD+4IL-17+ T-helper 17(Th17) cells,CD+8 IL-17+ T cells,CD+4 CD+25 FOXP+3 T regulatory (Treg) cells among activated T cells.Results Differentiation of Treg cells,Th17 cells and CD+8 IL-17+ T lymphocyte cells was enhanced when murine splenic T cells were cultured with TGF-β.The levels of expression were (7.8±2.2)%,(12.6±3.1)%,(10.1±2.6)% ,respectively.Experimental control group was severally same type of T cells without cytokine treatment.The levels of expression were (4.8±0.6) %,(1.7±0.5) %,(1.0±0.4) %,respectively.There were statistically significant differences among them (q=4.09,8.80,9.61.P<0.05 or P<0.01).Under combination treatment with IL-6 and TGF-β,(17.8±5.3) % Th17 cells and (15.0±4.2)% CDCD+8 IL-17CD+ T cells were induced,whereas the levels of Treg cells whose differentiation were restrained were (4.1±1.2) %.The differences were statistically significant compared with the level of same type of T cells in TGF-β group (q=5.03,5.17,5.04,P<0.01).Moreover,combination treatment with IL-2 and TGF-β decreased percentages of Th17 and CDCD+8 IL-17CD+ T cells and increased percentages of Treg cells in T cell population.There was an opposite effect when anti-IL-2 was apphed.The percentages of Th17 and CD+8 IL-17+ T cells were increased and the percentages of Treg cells were reduced The regulation trend of T lymphocyte cells in human peripheral blood was similar with those in routine spleen.Conclusion Various cytokines are of great importance in the regulation of the balance between Th17 and Treg cell.

Objective To investigate the clinical characteristics and its prognostic factors for hilar cholangiocarcinoma with exairesis .Methods The clinical data of 58 cases of hilar cholangiocarcinoma were ret-rospectively analyzed .The Kaplan-Meier method was used to estimate the overall survival and disease specific survival rates for these patients .And the factors that may influence the prognosis and survival of patients were an -alyzed using univariate(log-rank test)and multivariate Cox proportional hazard models .Results The median survival time was 22 months for all patients .The 1-,3-and 5-year survival rates were 76%,40%and 21%, respectively.Univariate analysis showed that preoperative albumin (P=0.002),intraoperative blood loss (P=0.039),surgical method(P =0.006),histologic differentiation(P =0.001),portal vein encroached(P =0.014),surgical margin(P=0.020)were correlated factors for postoperative survival duration .Multivariate analy-sis by Cox Proportional Hazard Model showed that surgical method (P=0.022),histologic differentiation(P=0.020)were independent prognostic factors for patients with Hilar cholangiocarcinoma excised .Conclusion Low albumin leve,intraoperative blood loss more than 500 mL,low degree of tumor differentiation,portal vein en-croached ,radical surgery ,positive surgical margin are risk factors for total survival of Hilar cholangiocarcinoma .

OBJECTIVE To investigate genes associated with the drug-resistance of ?-lactam antibiotics in Pseudomonas aeruginosa(PAE) isolated from clinical patients. METHODS ?-Lactamase genes including TEM,SHV,OXA-10,PER,VEB,GES,CARB,IMP,VIM,SPM,GIM,DHA,FOX,MOX and oprD2 were detected by PCR amplification in 33 PAE isolates. RESULTS TEM,SHV,GES,CARB and VIM genes were positive in 100%,6.1%,6.1%,9.1% and 12.1% of 33 isolates,respectively.The deletion of oprD2 gene was found in 22 isolates.Other ?-lactamase genes were absent in all isolates.By PCR amplification,DNA sequencing and BLASTn comparison analysis,the CARB genes of 2 strains were demonstrated to be CARB-3 and the VIM genes of 2 strains were VIM-2. CONCLUSIONS P.aeruginosa carries various beta-lactamase genes in clinical PAE patients,and the deletion ratio of oprD2 gene is high.

OBJECTIVE To investigate the existence of genes for beta-lactam antibiotic resistance and for aminoglycosides modification enzymes(AMEs) in Pseudomonas aeruginosa(PAE) isolates from ICU patients and analyze the homology among strains.METHODS ?-Lactamase genes including TEM,SHV,OXA-10,PER,VEB,GES,CARB,IMP,VIM,SPM,GIM,DHA,FOX,MOX and oprD2,were detected by PCR amplication in 21 PAE isolates.The genes for AMES including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰwere determined by PCR amplification as well.RESULTS Among 21 isolates 21(100%),2(9.5%),1(4.8%),2(9.5%)and 4(19.0%) were positive for TEM,SHV,GES,CARB and VIM genes,respectively.The deletion of oprD2 gene was found in 14 out of 21 strains.Other ?-lactamase genes were absent in all isolates.As for AME genes,aac(3)-Ⅱ,aac(6″)-Ⅰ,aac(6)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aac(3)-Ⅰgenes were present in 19.0%,23.8%,9.5%,4.8%,and 19.0% of 21 isolates,However,aac(3)-Ⅰ gene was no position in any isolates.CONCLUSIONS P.aeruginosa carries various beta-lactamase and AME genes in ICU patients.Genetic cluster analysis suggested that clonal propagation result in nosocomial infection of PAE.

The Acetobacter xylinum and Schizosacchromyces sp. were isolated from tea fungus culture. Each strain 's function on the production of antibacterial protein was explored by single-strain culture and double-strain culture under various condition. The preliminary conclusion is that the protein is synthesized by the yeast cells and its antibacterial activity is significantly improved through modification by bacteria enzymes. The results of co-cultures of several yeasts with the Acetobacter xylinum show that Schizosacchromyces is superior to other yeasts.

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.

AIM: To determine the effect of heat treatment on rat primary cultured neurons, and give fundamental research for candidate molecule to protect the neurons from heat injury. METHODS: Neurons from rat striatum were primary cultivated in D-MEM with 15% horse serum, and when got mature, cells were identified by immuno-cytochemical staining with neurofilament protein (NF), tyrosine hydroxylase (TH) and neuron specific enolase (NSE) antibodies. Cells in heat treatment groups were put in an 43 ℃ CO 2 incubator for 1 h, and the control groups at 37 ℃ as normal. Striatum neurons were stained with trypan blue and dual fluorscence dye (PI/H33258) immediately followed heat treatment, and necrosis rate of neurons was estimated. At the same time, activated caspase-3 immuno-cytochemical and TdT TUNEL methods were applied to determine apoptosis rate, and cell volume was also identified with micro-photography. RESULTS: During day 7 to day 9, the cultured striatum neurons got mature, and many neuronal fibers starched out and formed neuron network, NF, TH, and NSE staining positive. Treatment at 43 ℃ for 1 h, cell number decreased greatly, while NF+ percentage kept unchanged, and the heat treatment survived neurons were processing cell necrosis and apoptosis, but necrosis percentage was much greater than that of apoptosis. While cell volume kept unchanged after heat treatment. CONCLUSION: Heat treatment greatly affects the growth and survival of the cultured striatum neurons, and the injury effect is most due to cell necrosis process.

Objective To evaluate the effects of intrathecal methotrexate on the activation of microglia in spinal cord in a rat model of tibial cancer pain (TCP).Methods Thirty female Sprague-Dawley rats,aged 5-7 weeks,weighing 150-180 g,were randomly divided into 3 groups (n =10 each):sham operation + artificial cerebrospinal fluid (group SA),TCP + artificial cerebrospinal fluid (group CA),and TCP + methotrexate (group CM).TCP was induced by injecting Walker-256 cancer cells into the medullary cavity of tibia.Artificial cerebrospinal fluid or methotrexate 100μg (15μl) was injected intrathecally over 10 min on 7th day after TCP.Mechanical pain threshold (MPT) was measured before TCP,at 1,3,5 and 7 days after TCP and 2,4,8 and 24 h after administration (T0-8).The rats were sacrificed after measurement of the pain threshold at T8 and the spinal cord was isolated for detection of the activation of microglia (by immunofluorescence) and content of tumor necrosis factor-α (TNF-α) and IL-1β (by ELISA).Results Compared with group SA,MPT was significantly decreased,and the number of activated microglia cells in the spinal cord was increased,and the contents of TNF-α and IL-1β were increased in groups CA and CM (P ＜ 0.05).Compared with group CA,MPT was significantly increased,and the number of activated microglia cells in the spinal cord was decreased,and the contents of TNF-α and IL-1β were decreased in group CM (P ＜ 0.05).Conclusion The mechanism by which intrathecal methotrexate reduces TCP in rats is related to inhibition of the activation of microglia and reduction of the secretion of proinflammatory cytokines TNF-α and IL-1β in the spinal cord.