Objective To screen and identify protein interacted with enterovirus 71 (EV71)3A protein by means of T7-phage display system.Methods The prokaryotic expression vector of 3A protein was constructed for expressing and purifying 3A pro-tein.With 3A protein as the target protein,the T7-phage display system was adopted to screen the human hepatocyte cDNA library. The screened products were performed the DNA sequence analysis and the homologous research.Results 3A protein was expressed and purified,after 4 rounds of biopanning,37 positive clones were selected.The inserted fragments were amplified by PCR using specific T7 primer.The products were sequenced,2 proteins were identified to interact with 3A protein by homologous analysis. Conclusion Trough the T7 phage display system,two proteins interacted with 3A are obtained.The possible function of 3A protein can be speculated by researching the function of these two proteins,which lays the foundation for the further studying the pathogen-ic mechanism of EV71.

Objective To examine and analyze the methylation status of the promoter region of human telomerase reverse transcriptase(hTERT)in gastric cell lines.Methods Telomerase activity was detected with telomeric repeat amplification protocol(TRAP).Expression of hTERT was observed by immunofluorescence.The target sequence of hTERT CpG island(-692~-403bp)was selected by database search.The methylation status of hTERT CpG island was examined with bisulfite sequencing PCR(BSP)as following:Genomic DNAs were modified by sodium bisulfite and amplified by PCR with primers that contained no CpG sites.Direct sequencing was performed for each case.Results Both TRAP and immunofluorescence analysis showed that high levels of telomerase and hTERT were expressed in SGC-7901 and BGC-823 cells.The sequences(-634~-403bp)of hTERT gene promoter,which were sequenced successfully,were homologous to the sequences before sodium bisulfite modification.25 CpGs in BGC-823 cell line and 26 CpGs in SGC-7901 cell line were observed to be methylated in all 26 CpGs of this part.Only one repressive transcription factor,which called myeloid-specific zinc finger protein 2(MZF-2),was found to bind this region which possessed two special binding motifs.Such a kind of binding showed potential importance for methylation-mediated derepression of hTERT transcription.Conclusion The hypermethylation may change the structure of DNA and result in abortive binding between MZF-2 and its motif,which might be one of the reasons for up-regulation of hTERT transcription and telomerase activity.

Objective To investigate efficacy and effect on liver function of the experimental therapy with balloon catheter to block the main artery temporarily and then pressurize chemoembolization to treat hypovascular liver cancer. Methods Eighty patients with hypovascular liver cancer requiring interventional therapy were randomly divided into two groups.The experimental group was treated with the new therapy and the control group was treated with traditional therapy. The lipiodol-filling status and maximum diameter of the tumor was analyzed for a midterm outcome,and the change of AFP and liver function were evaluated.Mann-Whitney test was used for data between two groups,Friedman test was used for data of each group,and Spearman nonparameter relevant analysis was used for efficacy indexes.Results ( 1 ) All Patients have confirmed diagnosis of hypocvascular liver cancer and got balanced baseline. ( 2 ) Lipiodol-filling status:the clinical efficacy and benefit rates of patients from experimental group were higher than that from control group and showed statistically significant difference in 1,3,12 months (Z =-2.135,- 2.939,- 2.686 ; P =0.034,0.004,0.007 ),but no statistically significant difference in 6 month ( Z =- 1.170,P =0.242 ).The status of lipiodol-filling of experimental group ( x2 =2.593,P =0.459 ) was more stable than control group ( x2 =10.886,P =0.012).(3) Maximum diameter of the tumor:the clinical efficacy and benefit rates of patients from experimental group were higher than that from control group and showed statistically significant difference in 3,12 months ( Z =- 2.734,- 2.733 ; P =0.006,0.006),but no statistically significant difference in 1,6 month ( Z =- 1.692,- 1.895 ;P =0.091,0.058). But neither of two groups showed statistically significant difference in change of maximum diameter of the tumor ( x2 =5.500,P =0.139 ; x2 =6.509,P =0.089 ).Relation between lipiodol-filling and maximum diameter showed positive correlation in 3 month ( r =0.257,P =0.035 ). (4) Stratified analysis was used for data of AFP according to AFP value before therapy,and two groups showed no statistically significant difference for patients belonging to 20-1000 μg/L by Pearson Chi-square test. (5)Two groups showed no statistically significant difference for data of liver function before therapy and in 1,3,6months ( Z =- 1.073,- 1.314,-0.518,-0.549;P=0.308,0.189,0.604,0.583).Conclusions According to the midterm result of this experiment,the experimental therapy increased lipiodol-filling and decreased maximum diameter of the tumor significantly in 3 and 12 months correspondingly,but no significant difference was observed in AFP and liver function between groups yet. So the long-term efficacy and its influence to lung metastasis and survival rate need further research.

Moyamoya disease may cause subarachnoid hemorrhage because of its feature of spontaneous occlusion of the circle of Willis with vascular network abnormal hyperplasia on the base of the brain.This article reviews the disease characteristics of moyamoya disease-related subarachnoid hemorrhage,the correlations between bleeding mechanisms,disease progression and bleeding,as well as its therapeutic measures.

Objective The study aimed to compare the vector system with the hand instrument in the pain severity during scaling and root planning.Methods 60 periodontal patients were randomly divided into two groups for supportive periodontal therapy (SPT):the experimental group (using vector system) and the control group (using gracey instruments),with 30 cases in each group.And the painfulness after SPT was evaluated by Visual Analogue Scale (VAS).Results 66％ patients felt mild pain during the supportive periodontal therapy with the application of vector system,while 36％ patients felt mild pain in the control group.24 patients in the experimental group accepted vector system for SPT and 23 patients in the experimental group felt less fear of the treatment.Conclusions Patients will feel less discomfort with the application of the vector system,therefore better compliance will be reached.And the cooperation of the doctors and nurses has great impact on the treatment effect.

Objective To investigate the expression of Pin1 in human pancreatic carcinoma as well as adjacent tissues and to discuss the role of Pinl in oncogenesis of pancreatic carcinoma. Methods Specimen of pancreatic carcinoma tissues and adjacent tissues were collected from 20 cases. Pin1 mRNA and protein expression in pancreatic neoplasm and corresponding adjacent nontumorous tissues were detected by real-time quantitative reverse transcription-polymerase chain reaction (RQ-RT-PCR) and western blot. Results Pin1 was overexpressed at mRNA and protein level in pancreatic carcinoma tissues compared with that in their nontumorous counterparts ( 2.78 ± 1.02 vs 4.36 ± 1.27;5. 48 ± 1.69 vs 9.97 ± 1.86, P ＜ 0.05 ). Pin1 expression was not correlated to clinical stage and pathological grading of the carcinoma. Conclusion Pin1 overexpression may play a key role in pancreatic carcinoma.

OBJECTIVE: To investigate the clinical feature, diagnose and therapeutic methods of paraglottic space primary paraganglioma. METHOD: One case of paraglottic space primary paraganglioma was reported and the relevant literatures were reviewed. RESULT: One case showed a hoarse voice, who was cured after the surgery of neck incision. NSE and CgA were positively expressed. CONCLUSION Paraganglioma of the paraglottic space is very rare. The diagnosis of paraglottic space primary paraganglioma bases on histopathology and immunohistochemistry. The immunohistochemistry and clinical character must be comprehensively analyzed to increase the diagnosis accuracy.
Female ; Glottis ; pathology ; Humans ; Middle Aged ; Paraganglioma ; pathology ; surgery

Objective To establish and optimize AFLP reaction system used in providing necessary technique basis for genetic diversity analysis in Asarum sieboldii.Methods Leaves of A.sieboldii were used as experimental materials to analyze various essential elements of the whole process,such as quality of extracted DNA,time of restriction digest and ligation,concentration of Mg2+,primers and Taq DNA polymerase during PCR amplification etc.,so that the most optimal AFLP reaction system could be built up. Results The optimal AFLP reaction system of A.sieboldii has been constructed: in the genomic DNA extraction,mercaptoethanol being utilized and samples being incubated about 30 min at 65 ℃;the genomic DNA being digested by Trul Ⅰ and Pst Ⅰ for 3 h respectively;in PCR amplification,the final concentration of Mg2+ being 1.5 mmol/L and the volume of Taq DNA polymerase being 0.2 ?L.Conclusion The present reaction system for A.sieboldii is able to gain the favorable results of AFLP analysis and can be used in the genetic diversity research of the species.

BACKGROUND: It is difficult to culture human dental pulp cells in vitro. Studies regarding effects of growth factors on proliferation and differentiation of dental pulp cells cultured in vitro have been reported. However, little is known about the Chinese herb rhizoma drynariae decoction on dental pulp cells cultured in vitro.OBJECTIVE: To observe the effects of different concentrations of rhizoma drynariae decoction on the proliferation and differentiation of human dental pulp cells cultured in vitro.DESIGN, TIME AND SETTING: A controlled observation was performed at the Scientific Resaarch Center, Fourth Hospital, Hebei Medical University between March 2006 and May 2007.MATERIALS: Human dental pulp cells were sourced from the patients who acquired orthotherapy through pulling out impacted wisdom tooth at the Department of Stomatology, Fourth Hospital, Hebei Medical University. Written informed content of sample collection was obtained from all patients. Rhizoma drynariae (place of production: Yunnan Province in China) was provided by the Dispensary of Traditional Chinese Medicine, Fourth Hospital, Hebei Medical University.METHODS: Human dental pulp cells were cultured in vitro using method of tissue piece. The effective ingredients of rhizoma drynariae were extracted by alcohol deposition. 1 mL of physic liquor contained 1 g crude drug and diluted into 10, 50, 100, 500, and 1000 mg/L culture medium utilizing fetal bovine serum. Subsequently, the prepared culture medium was used to culture human dental pulp cells in vitro. Cells that were cultured using culture medium without rhizoma drynariae decoction were used as controls.MAIN OUTCOME MEASURES: ①Primary culture and source identification of human dental pulp cells. ②Effects of different concentrations of rhizoma drynariae decoction on proliferation of human dental pulp cells by methyl thiazolyl tetrazolium (MTT) assay. ③ Effects of different concentrations of rhizoma drynariae decoction on fibronectin expression in human dental pulp cells by immunohistochemistry. ④ Effects of rhizoma drynariae decoction on ultrastructure of human dental pulp cells utilizing scanning electron microscope and transmission electron microscope.RESULTS: Primarily cultured human dental pulp cells displayed polygon- and shuttle-shaped appearance. Different concentrations of rhizoma drynariae decoctions, in particular 100 mg/L, exhibited proliferation-promoting effects on proliferation of human dental pulp cells, and could induce dental pulp cell synthesis and secrete fibronectin. Electron microscopy results revealed that following treatment of rhizoma drynariae decoctions, human dental pulp cells were found with abundant ridges on their surface, surround by extracellular matrix, cytoplasm full of abundant rough endoplasmic reticulum and dissociative ribosome, as well as evenly dispersed nuclear euchromatin, and occasionally seen heterochromatin.CONCLUSION: 100 mg/L rhizoma drynadae decoction apparently promotes the proliferation of human dental pulp cells cultured in vitro.

Objective Shunaoxin Dropping Pills (SDPs), a Chinese patent medicine, has been used widely in China for the treatment of headache, amnesia, and insomnia. The aim of the present study is to observe the effect of SDPs on inducing angiogenesis and neurogenesis in vitro. Methods The present testing system using the serum obtained from animals ig treated with SDPs and a co-culture system in vitro was used to investigate if SDPs promotes brain microvascular endothelial cells (BMECs) tube formation and neural differentiation of neural stem/progenitor cells (NSPCs), which plays important roles in angiogenesis and neurogenesis. Results The SDPs serum sampled from rats ig treated with SDPs for 3 d dose-dependently promoted the tube like structure formation of cultured BMECs, and enhanced the fraction of MAP-2 positive cells of NSPCs, which co-cultured with the BMECs and astrocyte. In addition, there was no significant change in the percentage of glial fibrillary acidic protein positive cells. Conclusion Our results show that SDPs serum can induce neural differentiation and BMECs tube formation in vitro.