Objective:To investigate the effects of epidermal growth factor (EGF)on cell cycle and cell cycle-related regulatory factors of human esophageal squamous cell carcinoma (ESCC) cell line Eca109.Methods: Serum starved Eca109 cells were treated with 20 ng/ml recombinant human EGF(rhEGF)for 24 h.The cell cycle phase distribution was detected by flow cytometry.The mRNA and protein expression levels of p21CIP1/WAF1(p21) and p27KIP1(p27) were detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot,respectively.Results: The proportions of G1 phase cells in EGF group and control group were ( 54.90 ±0.82 )% and ( 65.94 ±0.74 )%.The mRNA and protein expression levels of p 21 in EGF group was significantly higher ,and p27 was significantly lower than that in control group ( P<0.01 ) .Conclusion: EGF facilitates G1-S phase transition,and promotes the proliferation of Eca 109 cells,which may be associated with the up-regulation of p21 and down-regulation of p27.

Objectives To explore the calcium signaling mechanism of STIM1 in breast cancer cells. Meth-ods After SiRNA interruption, Western blot and Transwell were used to measure protein expression of STIM1 and cell migration in MDA-MB-231 cells respectively. The relationship between STIM1 and SOCE calcium signaling were analysed by Laser confocal microscopy. Western blots were used to measure protein expression of FAK after si-lence STIM1. Results The numbers of cells without STIM1 were significantly lower than those cells with STIM1 by Transwell assay. STIM1 mediated SOCE in MDA-MB-231. Blocking SOCE might inhibite cells migration. Si-lence STIM1 did not affect the expression or activation of FAK in MDA-MB-231 cells. Conclusion STIM1 influ-ences cell migration through SOCE pathway in breast cancer cells, which is independent on the expression or activa-tion of FAK.

Objective To investigate the oropharyngeal microbial colonization in elderly patients.Methods Totally 618 elderly cases and 96 young and middle-aged cases were involved in this investigation.The colonization of bacteria and fungi on the oropharyngeal mucosa were obtained by throat swab cultures.Results There were 85.4％ of cases (82 cases) with Streptococcus viridans and Neisseria gonorrheae on the oropharynx mucosa in young and middle-aged group and the pattern of constitute was simple.3 to 5 bacterial species were isolated from the oropharyngeal mucosa in elderly group.There were only 25.7％ of cases (159 cases)with Streptococcus viridans and Neisseria gonorrheae on the oropharyngeal mucosa in elderly group.Biodiversity of the constitution pattern in the bacterial colonization was exhibited in the elderly.The colonization rate of Gramnegative bacilli was higher in elderly group than in young and middle-aged group [53.1％ (328/618)vs.6.3％ (6/96)].Staphylococcus aureus and Streptococcus pneumonia were the common Grampositive bacteria colonization in the elderly.The colonization rate of Candida fungus was 9.1 ％ (56/618) in the elderly.Conclusions The reduction of commensal bacteria,especially Streptococcus viridans may be the pathological basis of mode changes in bacteria colonization and opportunistic bacteria colonization on oropharyngeal mucosa in the elderly.The colonization rate of oropharyngeal Gram-negative bacili is obviously increased and Klebsiella Pneumoniae is the common bacteria on oropharyngeal mucosa in the elderly.Enterobacteriaceae,Pseudomonas aeruginosa and Acinetobacter baumannii are sensitive to common antibiotics.

Objective To explore the prognostic value of APACHE Ⅱscore in patients with Severe acute organophosphorus poi-soning .Methods 42 patients with Severe acute organophosphorus poisoning ,in which 34 cases survived ,8 cases dead ,were select-ed .The APACHE Ⅱscores of patients in first 24 h of admission were collected ,and receiver operating characteristic curves (ROC curve) were drawn .Results APACHE Ⅱ score of the 42 patients with Severe acute organophosphorus poisoning was 18 ~30 (20 .11 ± 6 .32) ,in which the survival group was(16 .10 ± 3 .12) ,the dead group was(28 .01 ± 4 .46) (P<0 .01) .With the increase of APACHE Ⅱ score ,the fatality rate gradually increased .The total area under the ROC curves of APACHE Ⅱ score for death judgment was 0 .922 ,APACHE Ⅱ score of 21 .2 was the best diagnostic point ,the sensitivity was 95% ,and specificity was 89% . Conclusion The APACHE Ⅱscore could predict severity of patients with Severe acute organophosphorus poisoning ,and APACHEⅡscore ≥21 .2 could be used as the prognosis for death of the patients .

Objective To clone and express the polymorphic membrane protein I(PmpI)gene of Chlamydia trachomatis(Ct), and to assess the immunogenicity and biological characteristics of PmpI. Methods A bioinformatic software was used to analyze the sequence of the PmpI gene of Ct, and to predict B cell epitopes in PmpI. With Ct serovar D DNA as the template, PCR was performed to amplify the N?terminal region(from position 90 to 1464)of the PmpI gene, which was cloned into a prokaryotic expression vector pET28a to express the recombinant protein PmpI. A Ni?ion affinity chromatography column was used to purify the recombinant protein, which was used to immunize New Zealand rabbits for preparation of polyclonal antibodies. Western blot analysis was conducted to evaluate the immunogenicity of this protein. Results A comprehensive analysis was carried out on the secondary structure, flexible regions, hydrophilicity plot, antigenic index and surface probability plot of the protein, which suggested that PmpI had 8 dominant B?cell epitopes. The product of PCR targeting the PmpI gene of Ct serovar D showed a total length of 1 375 bp. The recombinant prokaryotic expression vector pET28a?PmpI was successfully constructed. A recombi?nant protein with a relative molecular mass of approximately 50 000 was successfully expressed after isopropylβ?d?1?thiogalactopyranoside (IPTG) induction, and purified by affinity chromatography. Polyclonal antibodies against the recombinant protein were successfully prepared. Conclusion The N?PmpI protein of Ct serovar D is cloned and expressed successfully, laying a foundation for further studies on its biological functions.

Aliphatic amines in infant food packaging materials were extracted and concentrated by 0 . 5 mL of acidified methanol using gas purge microsyringe extraction ( GP-MSE ) . Pre-column fluorescence labeling of amines was achieved in mild conditions with 10-ethyl-acridine-2-sulfonyl chloride ( EASC ) as labeling reagent. The derivatization was carried out at 60℃ and pH 10. The derivatives were successfully separated on a Hypersil GOLD column with excitation and emission wavelengths of 262 and 430 nm, respectively. The detection limits were in the range of 0. 4-0. 6 μg/kg, and the quantitation limits were in the range of 1. 2-2. 1 μg/kg. All analytes were in good linearity in the concentration range of 2. 0-2000 μg/L with correlation coefficients of higher than 0. 998. The developed method was characterized by celerity, accuracy and high sensitivity. It was successfully applied to the determination of aliphatic amines in infant food packaging materials.

ObjectiveTo construct the eukaryotic expression plasmids containing cysteine protease inhibitor (CPI) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene from periodic Brugia malayi (Bm),and to observe its cellular immune response in mouse.Methods pcDNA3.1 (+)-BmCPI/BmGAPDH was constructed.The recombinant plasmids were screened and identified by digestion with restriction enzyme.BALB/c mice were injected intramuscularly with a dosage of 100 μg purified recombinant plasmid DNA with GpG oligodeoxynucleotide (CpG ODN) and two same doses were administrated at 2-week intervals.pcDNA3.1 (+) and phosphate buffered solution (PBS) were used as controls.The tissue of muscles at 4 weeks after the third injection was collected and the target gene was detected by reverse transcription-polymerase chain reaction (RTPCR).Two weeks after the third immunization,the stimulation index (SI) of spleen lymphocytes of immunized mice was measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method and the serum levels of interleukin (IL)-4 and interferon (IFN)-γ were detected by enzyme-linked immunosorbent assay (ELISA).The data were analyzed by t test.ResultsBmCPI/BmGAPDH gene in the injected muscle of the immunized mice was detected by RT-PCR. At 6 weeks after immunization,the SIot spleen T lymphocytes in pcDNA3.1 (+)-BmCPI/CpG group and pcDNA3.1 (+)-BmCPI/BmGAPDH/CpG group were 1.466 ± 0.635 and 1.610 ± 0.112,respectively,which were both higher than PBS group and pcDNA3.1( +)-CpG group (1.004 ± 0.019 and 1.078 ± 0.129,respectively) (t=64.438,45.318,42.749 and 34.314,respectively; all P＜0.05).At 4 weeks after immunization,the serum levels of IL-4 and IFN-γ of mice in pcDNA3.1 ( + )-BmCPI/BmGAPDH/ CpG group were significantly higher than those in pcDNA3.1 (+)-CpG group (t=288.053 and 76.453,respectively; both P＜0.05),while the serum level of IFN-γ was also higher than that in pcDNA3.1 (+)-BmCPI/CpG group (t=129.642,P＜0.05). ConclusionThe recombinant eukaryotic plasmid pcDNA3.1 (+)-BmCPI/BmGAPDH could be expressed in mice,and could elicit specific cellular immune responses in immunized mice.

Objective To observe the therapeutic effect of heat-clearing and dampness-removing therapy combined with Bifico for ulcerative colitis ( UC) induced by trinitrobenzene sulfonic acid ( TNBS), and to explore its influence on tumor necrosis factor alpha ( TNF-α) and interleukin-10 ( IL-10) in rats. Methods Forty female adult Sprague-Dawley rats were evenly randomized into five groups, namely normal control group, model group, Bifico (175 mg/kg) group, Chinese medicine group(enema with Changdiqing 3.6/kg and oral use of Changyanling Recipe l) , and Chinese medicine plus Bifico group. After treatment, the damage of colonic mucous membrane was evaluated, and expression levels of IL-10 and TNF-α in colonic mucosa were observed through immunohistochemical assay. Results The degree of colonic mucosal injury was severer and the inflammation was more obvious in the model group than those in the normal control groups ( P<0.01) , and the above changes were relieved to various degrees in the medication group ( P<0.01 compared with those in the model group) . Chinese medicine plus Bifico group had better effect on reducing colonic mucosa damage index ( CMDI) and inflammation, and on promoting the healing of colonic mucosa than Bifico group and Chinese medicine group (P<0.05 ) . The expression level of TNF-α in the colonic mucosa was markedly increased while that of IL-10 was markedly decreased in the model group ( P<0.01 compared with those in the normal control group) . The medication groups could counteract the above changes in the colonic mucosa ( P<0.01 compared with those in the model group) . The combination group and Chinese medicine group had better effect on decreasing TNF-α expression level and on increasing IL-10 expression level than Bifico group ( P<0.05) . Conclusion IL-10 and TNF-α play an important role in the pathogenesis and development of ulcerative colitis. Chinese medicine combined with Bifico has satisfactory therapeutic effect on UC rats, and its mechanism may be related with the increase of IL-10 expression level and with the decrease of TNF-α expression level.

Objective To discuss the importance of saving the life of patients and the severity assessment in early emergency for the trauma and hemorrhagic shock patients in the first-aid of pre-hospital and emergency department.Methods Retrospective anal-ysis of the 182 patients data.Results With early and aggressive treatment,177 cases of survival,5 cases of death,and the survival rate 97.3%.Conclusion The disease of traumatic shock patients is complex,high mortality.the early detection,taking timely and effective rescue measures are the key to increase the survival rate.

Objective To systematically review the effectiveness and safety of pantoprazole ( PAN ) vs. ranitidine (RAN) for patients with gastroesophageal reflux disease (GERD). Methods PubMed,Medline,EMbase,The Cochrane Library and three Chinese literature databases (CNKI,VIP and Wan fang) were retrieveed.Randomized controlled trials (RCTs) which compared the clinical outcomes of PAN group vs. RAN group for GERD were included. Two reviewers independently screened literatures in accordance with the inclusion and exclusion criteria, extracted the data and assessed the methodological quality of included studies.Then,meta-analysis was performed using RevMan 5.2 software. Results A total of 8 RCTs involving 1 590 patients were included.The results of meta-analysis showed that the PAN group was significantly superior to RAN group in terms of the healing rates and the relief rates of chief symptom for GERD of gradeⅠ-Ⅲ. While there was no significant difference in the incidence of adverse events between the two groups [GradeⅠ,RR=1.17,95%CI (0.80,1.70),P=0.43;GradeⅡorⅢ, RR=0.76,95%CI (0.43,1.36);P=0.36]. Conclusion Current evidence indicates that,pantoprazole is more effective than ranitidine for GERD of grade Ⅰ-Ⅲ,but both treatments are safe and well tolerated.