We wished to screen the cell line that stably expresses the HPV18E5 protein, and to ascertain the influence of HPV18E5 protein on cell proliferation and the cell cycle. The HPV18E5 gene was amplified by the polymerase chain reaction. Then, the His-tag pSecTag-HPV18E5 eukaryotic expression vector was constructed by digestion ligation and connection. The recombinant plasmid was transfected into Balb/c3T3 cells with lipofectamine, and positive cell lines were screened by a culture medium containing bleomycin. HPV18E5 expression in cells was confirmed by western blotting and immuno-enzymatic methods. The influence of HPV18E5 on cell proliferation and the cell cycle were detected by Cell Counting Kit-8 and flow cytometry, respectively. The pSecTag-HPV18E5 eukaryotic expression vector was constructed. After 21-day selection in a culture medium containing 400 μg/mL bleomycin, stably expressing HPV18E5 protein cells were harvested. Compared with control groups, cell proliferation in HPV18E5 stably expressed cells was obviously increased, as was the S phase in the cell cycle. Our results suggested that HPV18E5 influences cell proliferation and the cell cycle. Our study has laid the foundation of the biologic properties of HPV18E5 protein, which will aid further studies on the mechanism of action of carcinogenesis.
Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Human papillomavirus 18 ; genetics ; metabolism ; Humans ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomavirus Infections ; physiopathology ; virology ; Transfection

Objective To detect the genomic copy number variations(CNVs) of mice embryonic neural tissue with neural tube defects (NTDs) induced by methotrexate (MTX),and investigate the molecular genetic mechanisms between folic acid metabolism disorders and NTDs pathogenesis.Methods C57BL/6J NTD mice model was induced by MTX on gestational day 7.5,and the maternal serum and NTD embryonic neural tissues were collected;the array-comparative genomic hybridization(array-CGH) assay was utilized to analyze the whole genomic CNVs in NTD embryonic neural tissues;reverse transcription polymerase chain reaction(RT-PCR) was used to confirm the positive results;the maternal serum concentrations of folic acid and related metabolites and dihydrofolate reductase (DHFR) activity were detected by liquid chromatography-tandem mass spectrometry and enzymatic methods,respectively.Results ArrayCGH and RT-PCR results showed the 3 high confidence CNVs on XqE3,XqA1.1-qA2 and XqA1.1 in the NTD embryonic neural tissues.The NTD maternal serum concentrations of 5-methyltetrahydrofolate,5-formyltetrahydrofolic acid,S-adenosyl methionine and DHFR activity were reduced significantly compared with the control group,and there were statistical differences(all P ＜0.05).Conclusions There are obvious CNVs in embryonic neural tissue of NTD mice induced by MTX and folic acid dysmetabolism might cause mice embryonic neural tube developmental disorders through CNVs.

Objective To studythe cephalocaudal relationship ofabdominal aortic bifurcation relative toumbilicus and iliac crest vertex and their correlations with abdominal adipose tissue thickness and age. Methods The vertical distances,cephalocaudal relationship and other related anatomic parameters of aortic bifurcation relative to umbilicus and iliac crest vertex in 108 patientswere measured by consecutive abdominal CT scanning. The correlations of the acquired data with abdominal adipose tissue thickness and age were analyzed using Pearson correlation coefficient. Results Umbilicus was located at cephalad to aortic bifurcation in 67 patients(62.0%), caudal in 34(31.4%)andthe same level in 7(6.5%),with the vertical distance of(4.53 ± 17.51)mm to the aortic bifurcation. No statistically significant relationship was found between abdominal adipose tissue thickness(P>0.05) or age(P>0.05). Iliac crest vertex relative to aortic bifurcationwas located at cephalad,caudal and the same level in 31,71 and 6 patients,taking up 28.7%,65.7%and 5.6%,respectively. Its vertical distance to the bifurcation was(-6.34 ± 14.49)mm,nonrelated with abdominal adipose tissue thickness(P>0.05),but positively correlated with age(P<0.01). The difference in the cephalocaudal relationship of aortic bifurcation relative to umbilicus and iliac crest vertex was statistically significant(P<0.01). Conclusion Compared with iliac crest vertex,umbilicus is an important landmark of locating abdominal aortic terminal occlusion position in vitro because it mostly lies cephalad to aortic bifurcation in the front of the body,not easy to vary with abdominal adipose tissue thickness and age.

OBJECTIVETo study the characteristics of diagnosis and surgical treatment of esophageal leiomyoma.

METHODSPreoperative radiology, esophagoscopy and results of surgical treatment were retrospectively analyzed in 141 patients with esophageal leiomyoma.

RESULTSDysphagia (64.5%) and chest or epigastric pain (20.6%) were the most common presenting symptoms with a mean duration of 15.7 months. Eleven patients had been misdiagnosed as malignant esophageal disease, mediastinal or retroperitoneal tumor before operation. Thirteen patents received partial esophageal resection, including 2 patients with gigantic tumor, 3 still misdiagnosed as malignancy in the operation theater and 8 accompanied with esophageal or gastric cardiac carcinoma. 128 patients were treated with tumor removal only, among whom the esophageal mucosa was ruptured and sutured in 13 patents. The tumor was single in 130 patients and multiple in 11 patients. The tumor shape was either oval, horseshoe-or ginger-like. Tumor surrounded the esophagus less than half circumference in 75 patients, more than half in 59 patients and involved the whole circumference in 7 patients. The tumor was polypoid and intraluminal in one patient. It was extraluminal in 2 patients and was intramural in all the others. The morbidity and 30-day mortality rates were 3.6% and 0%.

CONCLUSIONPreoperative differentiation of esophageal leiomyoma is important for it may be misdiagnosed as esophageal malignant disease or mediastinal tumor. Tumor removal only is indicated if possible. Biopsy is inadvisable if leiomyoma has been suspected by esophagoscopy. The incidence of esophageal carcinoma complicating esophageal leiomyoma is high, of which the reason needs further study.

Adolescent ; Adult ; Aged ; Esophageal Neoplasms ; diagnosis ; surgery ; Female ; Humans ; Leiomyoma ; diagnosis ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Objective To construct the eukaryotic expression vector of human papillomavirus type18E5 and EGFP fusion gene,to analyze E5 expression in BALB/c3T3 cell.Methods The HPV18E5 and EGFP gene were amplified by polymerase chain reaction,and pSecTag-HPV18 E5-EGFP fusion gene eukaryotic expression vector was constructed by the method of digestion ligation and transformation.The recombinant plasmid was transfected into BALB/c3T3 cells with lipofectamine and the expressed product was detected by RT-PCR,the aim gene expression was observed under fluorescence microscope.Results The success in construction of pSecTag-HPV18 E5-EGFP fusion gene eukaryotic expression vector,RT-PCR result showed that HPV18E5 gene could be expressed in BALB/c 3T3 cell,fluorescent light could be observed under fluorescence microscope.Conclusion pSecTag-HPV18 E5-EGFP fusion gene eukaryotic expression vector has been successfully constructed,the recombinant plasmid could be expressed in BALB/ c3T3 cell.