AIM:To discuss Daidzein intravitreal injection whether has protective and recovery effects on acute nerve damages.
METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group ( 10μmol/L, 100μmol/L, 1000μmol/L ) and positive control group using rats nerve growth factor ( mNGF, 100ng/mL ). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups.
RESULTS: Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance (P<0. 05).CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.

Objective To screen the specific antigen mimotopes of influenza A(H1N1)virus by phage display technology,in order to pave a way to develop novel influenza vaccine.Methods Using human convalescent serum of pandemic influenza A(H1N1)in 2009 as solid-phase selective molecule,an artificial synthesized phage randomly displaying cyclic 7-mer peptide was screened with three rounds of "absorption-elution-amplification" selection.At the end of the third round selection,32 clones were randomly chosen from the top-agar phage plaques and placed onto E.coli ER2378 in logarithmic growth phase.After culturing for 5 hours,the positive clones were identified by enzyme linked immunosorbent assay(ELISA),cross reaction test and competitive inhibition assay.The identified positive clones were analyzed by DNA sequencing.The decoded amino acids sequences,which displayed on the surface of phage,were aligned with the hemagglutinin(HA)gene of influenza virus by homology comparison for definition of the mimotopes of influenza A(H1N1)virus.Results After enriching the specific antibody-binding phages from phage displaying peptide library,12 positive clones were chosen from 32 randomly selected clones.DNA sequencing and homology comparison showed that one epitope PLHARLP was confirmed as mimotope of swine influenza A(H1N1)virus antigen,which was composed of the 52nd,53rd,59th,60th,61st,83rd and 271st amino acid.Conclusions Swine influenza A(H1N1)mimotope has been obtained by cyclic 7-mer peptide phage library screening.This result provides a basis for developing new influenza virus vaccine from virus antigen mimotopes.

Objective:To express the B subunit of Shiga-like toxin type Ⅱ,and analyze its expression form and receptor-binding activity.Methods:The slt2b gene was obtained from EHEC O157∶H7 by PCR,and cloned to the expression vector pET22b(+).The genetically engineered bacteria pET22b(+)-stx2B/BL21 expressed the recombinant StxB after induced with IPTG.The renatured inclusion bodies were purified by ion exchange chromatography.The expression form of rStx2B was investigated by denaturing and native electrophoresis.The receptor-binding activity was confirmed by fluorescence detection and flow cytometer.Result:The constructed genetically engineered bacteria expressed the rStx2B at a high level.The purified protein was obtained after denaturation,renaturation and ion exchange chromatography.According to the denaturing and native electrophoresis,the rStx2B was expressed in a dimmer form,which consists of two monomers cross linked with disulfide bridge.The rStx2B showed good receptor-binding activity by Hela-binding assay.Conclusion:The genetically engineered bacteria were constructed successfully.The receptor-binding activity of rStx2B was independent of the pentamers.

Objective To investigate of the effect and mechanisman of SERCA2 on the phenotype modulation of HASMCs. Methods HASMCs were starved for 5 days and divided into different groups ,then we observed morphology change of the cells from the microscope and detected a-actin、SERCA2 and p-ERK by Western Blot,cells proliferation was observed by CCK-8 method. Results Compared with the control group,PDGF could reduce a-actin of HASMCs and increased the cells proliferation ability ,TSG could significantly inhibit the effect (P<0.01), PDGF could also significantly inhibit SERCA2 protein and increased the expression p-ERK (P<0.01), while U0126 significantly inhibited the effect (P < 0.01). Conclusion PDGF may induce HASMCs phenotype modulation through the regulation of SERCA2 and p-ERK.

Aim The expression of NF-?B activation and the effect of antioxidant (N- acetylcysteine, NAC) on NF-?B activity in human airway epithelial cell was assessed. Methods Using the TNF-?,the airway epithelial cell strains of normal subject(16HBE) and tumor patient (H292) was activated and using Western-Blot and ELISA the expression of NF-?B and IL-8 were detected. Results It was found that the activity of NF-?B could be stimulated by the TNF-? and increase with the amount of TNF-? with the peak occurring at 2 to 4 hours after stimulation and then decreasing at six hours. At the same time, the level of IL-8 was elevated, but decreased with inhibition of NF-?B activity by NAC, that means the action of NAC has a dose-dependent effect. Conclusion NAC not only blocks the signal transmission activated by NF-?B, but also anticipates the transcription modulation of expression of many cell factors and inflammatory mediums. It suggests that NAC may play a role in the anti-inflammatory treatment of respiratory diseases.

Antigens, CD20 ; metabolism ; CD5 Antigens ; metabolism ; Colonic Neoplasms ; complications ; metabolism ; pathology ; surgery ; Cyclin D1 ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Ileal Diseases ; complications ; pathology ; surgery ; Ileocecal Valve ; Intestinal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Intestinal Polyps ; complications ; metabolism ; pathology ; surgery ; Intussusception ; complications ; pathology ; surgery ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; complications ; metabolism ; pathology ; surgery ; Middle Aged

Objective To investigate the antibacterial effect of grape peel residue procyanidins on common clinical bacteria ,inclu‐ding E .coli ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,Enterococcus and methicillin‐resistant Staphylococcus aureus (M R‐SA) .Methods By the means of agar plate method ,we detected the minimum inhibitory concentration (MIC) of 30 strains of E .co‐li ,28 strains of Pseudomonas aeruginosa ,15 strains of Acinetobacter baumannii ,32 strains of Feces Enterococcus ,25 strains of E . faecalis and 70 strains of MRSA ,then calculate the MIC50 and MIC90 .The result was analyzed with SPSS software 16 .0 .Results The concentration of procyanidins at 3 -8 mg/mL had no inhibitory effect on E .coli ,Pseudomonas aeruginosa and Acinetobacter baumannii;there was no inhibitory effect on E .faecalis at 3-8 mg/mL ,but the inhibition rate to Feces Enterococcus was 6 .3% .By measuring the inhibitory effect of procyanidins on 70 MRSA ,the inhibition rate at 4 mg/mL was 65 .7% ,at 8 mg/mL was 95 .7% , MIC50 was 4 .221 mg/mL and MIC90 was 6 .260 mg/mL .Conclusion There are no inhibitory effects of grape peel residue procya‐nidins on Gram‐negative bacilli such as E .coli ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,and there are certain inhibitory effects on Gram‐positive bacteria such as MRSA ,enterococci ,especially on MRSA (P<0 .05) .

Objective To investigate the advantages of detection for EGFR gene mutations by denaturing high performance liquid chromatography (DHPLC) technology. Methods DHPLC was used to detect EGFR gene mutations at exon 19 and 21 in 49 cases of non-small cell lung cancer (NSCLC) patients,and the direct DNA sequencing was used to verify the accuracy of DHPLC detection. Results EGFR gene mutation was identified from 13 of 49 cases by DHPLC,including deletion mutation at exon 19 in 10 cases (76.92 %) and alternative mutations at exon 21 in 3 cases (23.08 %). Mutation results of DHPLC was consistent with DNA direct sequencing. The results of the direct DNA sequencing were the same as those of DHPLC. The sensitivity of mutation test by DHPLC was 100 %. Conclusion DHPLC technology can be used for large scale screening of EGFR gene mutation with rapid and accuracy.

The study aims to identify the role of cAMP-PKA pathway in the group Ⅱ metabotropic glutamate receptors (mGluRs)-mediated regulation of respiratory rhythm from the brainstem slice. Neonatal (aged 0-3 d) Sprague-Dawley rats of either sex were used. The brainstem slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared, and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O2 and 5% CO2) bubbling, and ended in 3 min. Respiratory rhythmical discharge activity (RRDA) of the hypoglossal nerve rootlets was recorded by suction electrode. Eighteen brainstem slice preparations were divided into 3 groups. In group 1, group Ⅱ mGluRs specific antagonist (2S)-α-ethylglutamic acid (EGLU) was added into the perfusion solution for 10 min. In group 2, after application of Forskolin for 10 min, washout with MKS, the slice was perfused with Rp-cyclic 3', 5'-hydrogen phosphorothioate adenosine triethylammonium salt (Rp-cAMPS) alone for another 10 min. In group 3, after application of Rp-cAMPS for 10 min, additional EGLU was added into the perfusion for another 10 min. The results showed EGLU shortened respiratory cycle (RC), but the changes of integral amplitude (IA) and inspiratory time (TI) were not statistically significant. Forskolin induced significant decreases in RC, and increased TI, IA. Rp-cAMPS could make the opposite effect compared with the changes of RRDA with Forskolin. The effect of EGLU on the RRDA was inhibited after blocking the cAMP-PKA pathway. Taken together, cAMP-PKA pathway may play an important role in the group Ⅱ mGluRs-mediated regulation of RRDA in the brainstem slice of neonatal rats.
Animals ; Animals, Newborn ; Brain Stem ; physiology ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Female ; In Vitro Techniques ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Metabotropic Glutamate ; physiology ; Respiration ; Signal Transduction ; physiology

Objective To compare the application and clinical significance of the liquid based cytology examination and the DNA quantitative analysis in female cervical lesions.Methods The cervical cell samples were collected from 879 women participating in the comparison by the cervical brush and performed the the liquid-based thin layer section preparation for conducting Papanicolaou staining and DNA staining respectively.The liquid based cytology examination was performed on the Papanicolaou staining section and the fully automatic scanning diagnosis was performed on the DNA staining section.Results The cases of above atypical squa-mous cells of undetermined significance(ASCUS)detected by the liquid based cytology examination and the partial cases of hetero-ploid cell detected by the fully automated DNA ploidy analysis system were recommended to further perform colposcopy and cervi-cal biopsy.28 women were performed the pathological biopsy.With the cytological examination result as the standard,the detection rate of above ASCUS cervical lesions detected by the cellular DNA quantitative analysis was calculated.Conclusion The combined application of the cellular DNA quantitative analysis method and the liquid based cytology examination can obviously increase the positive detection rate of cervical cancer and precancerous lesion,which has important significance for the prevention and treatment of female cervical cancer in our country.