10.3969/j.issn.1007-3969.2013.05.004

Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; therapeutic use ; Drug Resistance, Neoplasm ; Exons ; Gastrectomy ; methods ; Gastrointestinal Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Gastrointestinal Stromal Tumors ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Imatinib Mesylate ; Liver Neoplasms ; drug therapy ; secondary ; Male ; Piperazines ; therapeutic use ; Point Mutation ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Pyrimidines ; therapeutic use

Objective To express EV71 VP1 in prokaryotic expression system,initially evaluate the ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody.Methods Construct the prokaryotic expression plasmid pET30a (+)-VP1.Induced expression in Transetta (DE3) with IPTG and identified by Western blot.After purified with HisBind Protein Purification Kit,its ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody were analyzed.Results Plasmid PET-30a(+)-VPI was constructed successfully and the objective protein was expressed effectively.The ELISA titer of the polyclonal antibody was 1:3200 while neutralizing titer was 1:16 and the recombination protein lost the ability of blocking EV71 infection.Conclusion The recombination protein can stimulate mice to produce antibodies and the polyclonal antibody shew neutralizing activity but the recombination protein lost the binding activity to receptors probably due to the wrong advanced structure.

Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells .Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR.Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L.Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group , PBA group and negative control cells and nearly consistent among three groups , and as high as normal cells in combined treatment group . Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2 ’-deoxycytidine or 4-phenylbutyric acid separately , while could restore normal when treated with both agents , indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation .

Objective To study the application of Cell Counting Kit-8(CCK-8) in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cell .Methods The proliferation of K562 cells was detected by CCK-8 with different concentrations of 5-Aza-2 ’-deoxycytidine and the cell cycle and apoptosis of K 562 cells were detected after K562 treated by 50% inhibitory concentration of 5-Aza-2 ’-deoxycytidine .Results The 50% inhibitory concentration of 5-Aza-2’-deoxycytidine was 15.55nmol/L, after treated with this concentration , K562 cells showed that G2 phase arrest occurred , proliferation inhibited and apoptosis peaks appeared .Conclusion Inhibition of proliferation of K562 cells with different concentrations of 5-Aza-2’-deoxycytidine varied in a dose-dependent relationship , and 5-Aza-2’-deoxycytidine could promote apoptosis of K 562 cells.CCK-8 can be used in detecting the effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells .

AIM: (1) To investigate the effect of amniotic membrane transplantation (AMT) on rabbit conjunctival surface reconstruction with severe alkali burns. (2) To evaluate the possibility of AMT treatment for ocular alkali burns during recovering stage.METHODS: Animal models were established on 30 eyes of rabbits by creating severe alkali burns on the conjunctiva from the upper corneal limbus to the upper conjunctival fornix.Preserved human amniotic membrane transplantations and reconstruction of conjunctival fornix were performed at one week after injury (recovering stage). Epithelium growth of burned area after transplantation was observed using light microscope at 1, 2, 3, 4, and 8 weeks. Conjunctival tissue in transplantation area was collected at 1, 4 and 8 weeks. The ultrastructure of the collected tissue was studied by electron microscope. The results were compared with control group,which received only vitamin C subconjunctival injection and antibiotic eye drops as treatment for alkali burn. Exterior eye pictures were also taken at the end of the observation, the width from upper corneal limbus to the edge of upper fornix was measured. Data was analyzed statistically.RESULTS: 1) Tn the transplant group, conjunctival epithelium growth was observed in the area of AMT under both light and electron microscope 1 week after surgery. At 4weeks, conjunctival epithelium with goblet cells that resembled normal conjunctival tissues was observed in the whole amniotic membrane area. At 12 weeks, the conjunctival epithelium on the amniotic membrane was well formed, and the connective tissue under the epithelium was loose at the fornix. No fibrosis was identified. In contrast, conjunctival epithelium necrosis was observed in the control group at 2weeks after alkali burns. Re-epithelization did not occur through the 12-week observation. Severe fibrosis with inflammatory cells infiltration was observed between 4 to 8weeks. At 12 weeks, fibrosis of the connective tissue at the fornix developed and there were no conjunctival epithelium covering the burned area. 2) In the transplant group, the conjunctiva in transplanted area had no scarring and appeared smooth at 12 weeks. Upper fornix was reconstructed. The depth of fornix was 7.9±0.3mm (7.6-8.2mm), which was approximate to the normal depth 8.2±0.2mm (8.0-8.4 mm,P＞.05). While in the control group, the burned area appeared rough with granuloma formation and severe scarring. Upper fornix became shallow. The depth of fornix was 3.1±1.7mm(1.0 to 4.5mm.), and significant difference was found between control and transplant group (P＜0.01).CONCLUSION: Human amniotic membrane preserved in glycerin can promote cell adhering, migrating and differentiating of normal conjunctival epithelium.Reconstruction of conjunctival surface in early stage of alkali burn can be achieved by AMT. AMT can effectively prevent symblepharon formation.

Objective To appraise the bowel control and quality of life after transanal improvement of Soave one-stage pull-through operation for Hirschsprung′s disease(HD) in infants.Methods Twenty-five patients(aged from 2 months to 3 years old) with HD underwent improved endorectal pull-through procedure and postoperative follow-up for 5 months to 6 years(average 4.5 years) including the occurrence of operation related complications sush as bowel control,constipation or postoperative enterocolitis and quality of life.Results Twenty-one patients completed the study were with a mean follow-up.The assessment of anal continence was graded as normal(4 scores),good(3 scores),and fair(2 scores) according to the previous study of other investigators.Of the 21 patients,16 were graded as normal,whereas 4 were as good and 1 fair.One patient developed constipation and 2 patients had postoperative enterocolitis.Conclusion Modified Soave procedure for Hirschsprang′s disease can get a good bowel control outcome and quality of life.

Objective:To analyze the result of smooth pursuit test(SPT) in unilateral vestibular peripheral vertigo and investigate its influencing factors.Method:Smooth pursuit test (SPT) and spontaneous nystagmus (SN)were examined in one hundred and eighty-five patients with unilateral peripheral vertigo(case group) and 51 normal persons(control group) by Video-Nystagmography (Synapsys, France), and the gain of SPT and SN were selected as the observation parameters in order to analyze the waveform and gain of SPT and the relativity between SN and the gain of SPT.Result:Of the 185 patients, 105(56.8%),72(38.9%) and 8(4.3%) cases producedⅠ,Ⅱ and Ⅲ waveforms respectively. Of these patients, 58(31.4%) demonstrated SN and none had Ⅳ waveform. While of 51 normal persons, 38(74.5%), 13(25.5%) persons producedⅠand Ⅱwaveforms repectively and there were no Ⅲ, Ⅳ waveforms or SN. There was statistical significance between the stong and weak gain of SPT in these two groups. Weak gain was significantly different between two groups. The stong and weak gain of SPT in case group were 0.86±0.06,0.80±0.06; 0.78±0.09, 0.65±0.1; 0.68±0.13, 0.45±0.12. The relativity between SN and the gain of SPT was positive when they had same direction(r_s=-0.63,P＜0.05)and negative when opposite (r_s=0.34,P＜0.05).Conclusion:Ⅰ,Ⅱ,Ⅲ three waveforms of SPT could appear in unilateral vestibular peripheral vertigo and the corresponding gains are gradually decreasing.SN is the influencing factor of SPT.

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At present, a large number of studies have proved the important role of the nerve-gut-brain axis in mental disorders, and focused on the correlation between gut microbiota and mental disorders and the potential regulating mechanisms. This paper summarizes the current research status of gut microflora in several mental disorders, such as depression, anxiety and substance use disorders, in order to provide more clinical evidence for gut microflora as a new target for treatment.