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 investigate the effectiveness of intravenous thrombolytic therapy mode led by fast-track specialist nurses on patients with acute ischemic stroke(AIS).Methods This study involved 124 AIS patients who underwent intravenous thrombolytic therapy in the Department of Emergency of our hospital from March 2021 to February 2023.Among the patients,61 admitted between March 2021 and February 2022 received conventional AIS thrombolytic therapy were assigned to a control group.While the 63 patients who received AIS thrombolytic therapy under the specialist nurse-led intravenous thrombolytic therapy mode between March 2022 and February 2023 were assigned to an observation group.The two groups were compared in terms of the time from admission to completion of CT examination,time for signing the informed consent for thrombolytic therapy,door to needle time and percentage of DTN<60 minutes,as well as the post-thrombolysis scores according to the National Institute of Health Stroke Scale(NIHSS)and satisfaction to medical consultation.Results The observation group exhibited a significantly shorter time from admission to completion of CT examination,a shorter time for signing an informed consent for thrombolytic therapy,a shorter door to needle time and a higher percentage of DTN<60 minutes,all with significant difference in comparison with those in the control group.After thrombolysis,the NIHSS score of the observation group decreased more than that of the control group(P<0.05).The patients and their families in the observation group reported significantly higher satisfaction compared to those in the control group(both P<0.05).Conclusion The fast-track specialist nurse-led intravenous thrombolytic therapy mode demonstrates the superiority in reduction of the time from admission to completion of CT examination,time for signing an informed consent for thrombolytic therapy,door to needle time and the NIHSS scores,higher percentage of DTN<60 minutes as well as improvement of patient satisfaction.

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.

Nonalcoholic fatty liver disease （NAFLD） has become the most common chronic liver disease in the world， and it is also one of the main causes of liver cirrhosis and hepatocellular carcinoma， so it is particularly important to curb the development and progression of NAFLD in a timely manner. However， due to its complex pathogeneses， there are currently no effective methods for radical treatment. As a new generation of probiotics， Akkermansia muciniphila （Akk bacteria） can improve metabolic disorders of the body， and more and more studies have shown that Akk bacteria have a potential therapeutic effect on metabolic diseases， especially NAFLD. Therefore， this article briefly reviews the mechanism of action of Akk bacteria in NAFLD， in order to provide new ideas for improving the treatment of NAFLD and creating new therapies.