PAC1 is the neuropeptide pituitary adenylate cyclase activating polypeptide (PACAP) preferring receptor, which belongs to class B G protein-coupled receptors (GPCR) family. PAC1 mediates the most effects of PACAP as neurotransmitter, neuroregulator and neuroprotectant, while its high expression has close relationship with some physiological and pathological processes such as nerve-injury and tumor. To further understand the function of PAC1, a cell line that expressed inducible PAC1 was constructed to achieve Doxycycline (Dox) dependent expression of PAC1 in CHO (Chinese hamster ovary) cell using the improved Tet (tetracycline)-on Advanced System. First, the PAC1-EYFP fusion gene composed of PAC1 gene and gene encoding EYFP (enhanced yellow fluorescent protein) was sub-cloned to the tetracycline response element pTRE-Tight vector to construct the recombinant vector pEYFP-PAC1-EYFP by double enzyme digestion. Second, the tetracycline regulation components pTet-On advanced vector and the response element pTRE-PAC1-EYFP vector were both introduced into CHO cells successively and the positive clones were screened with G418 and hygromycin respectively. Third, the controlled expression of PAC1-EYFP in CHO was induced by tetracycline analogues Dox in different concentrations and the different levels of receptor PAC1-EYFP were detected. The results of fluorescence analysis and western blotting show that the cell strain with Dox dependent expression of PAC1-EYFP named PAC1-Tet-CHO was obtained. Moreover, in PAC1-Tet-CHO cells the expression of PAC1-EYFP was induced by Dox in a dose-dependent manner. The inducible expression of PAC1 still was stable after sub-culturing for more than 10 passages. It was also found by MTT assay that the higher expression level of PAC1 endowed the cells with higher proliferative viabilities. The construction of controlled expression system of PAC1 will lay a foundation for the further research on PAC1 profiles.
Animals ; Blotting, Western ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Genetic Vectors ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I ; biosynthesis

OBJECTIVE To analyze the risk factors,clinical characteristics and prevention countermeasures of(nosocomial) fungal infection in patients with acute leukemia.METHODS To adopt investigation way to review and analyze the clinical data of nosocomial fungal infection in 39 cases from 178 acute leukemia inpatients.RESULTS The(nosocomial) fungal infection rate was 21.91%.The nosocomial fungal infection rate in elderly group was(higher) than that of younger group,and in granulocytosis group(≥0.5?10~9/L) was higher than that of

Objective To observe the clinical efficacy of acupoint application withChan Wu gel cream plus five notes therapy in treating cancer pain.Method Eighty patients with cancer pain were randomized into a treatment group and a control group, 40 cases in each group. The treatment group received acupoint application withChanWu gel cream at the corresponding acupoints of the affected organ in association with the five notes therapy; the control group was intervened by the three-step analgesic ladder recommended by World Health Organization (WHO). The quality of life and pain intensity were scored before and after the intervention.Result The quality of life and pain degree were significantly changed after the treatment in the two groups (P<0.01). The quality of life and pain intensity of the treatment group were significantly different from that of the control group (P<0.05).Conclusion Acupoint application withChan Wu gel cream plus five notes therapy is effective in mitigating cancer pain and improving the quality of life.

Objective To study the application value of endoscopic purse-string sutures with titanium clips and endoloops for the gastric wall defect during endoscopic full-thickness resection (EFTR). Methods Data of iffteen hospitalized patients with gastric submucosal tumor (SMT) undergone EFTR was reviewed. The patients were all applied with endoscopic purse-string sutures with titanium clips and endoloops, which was performed after the EFTR when the gastric walls were perforated artiifcially. Results The gastric tumors were complete successfully resected in the iffteen patients through endoscopic surgery, applied the purse-string sutures with titanium clips and endoloops after the EFTR when the gastric walls were perforated artiifcially. Approximately, ifve clips were utilized on average. Postoperative surgical wound healing was followed up for 6 months, no tumors recurred. Conclusion It is safe, minimally invasive treatment method for rapid rehabilitation in endoscopic full-thickness resection utilized purse-string sutures with titanium clips and endoloops.

Objective:To investigate the long-term death of patients with ischemic stroke and its influencing factors.Methods:Based on the data of patients with ischemic stroke in the multi-center oral fibrinogen-lowering drug secondary prevention database, the follow-up patient information and the cause of death were registered through the epidemiological investigation method, and then compared with the baseline data of patients in the original database.Results:A total of 278 patients completed the follow-up, and 166 were in lumbrokinase group and 112 were in control group. There were 124 deaths (44.6%) within 10 years, of which 92 (74.2%) were vascular deaths. In the lumbrokinase group, 74 patients (44.6%) died of all causes and 55 (33.1%) died of vascular diseases; in the control group, 50 (44.6%) died of all causes and 37 (33.0%) died of vascular diseases. Cox proportional risk model analysis showed that lumbrokinase treatment had no significant effect on the 10-year survival rate of patients with ischemic stroke. The analysis of death influencing factors showed that the baseline international normalized ratio (INR) was significantly associated with the 10-year non-vascular death risk of patients (hazard ratio [ HR] 1.98, 95% confidence interval [ CI] 1.21-3.25; P=0.006). The greater the decrease of tissue plasminogen activator (tPA) within half a year, the lower the 10-year all-cause mortality risk ( HR 0.94, 95% CI 0.90-0.99; P=0.011); the greater the decrease in INR within one year , the lower the 10-year vascular death risk ( HR 0.41, 95% CI 0.17-0.96; P=0.040); the greater the decrease of D-dimer within one year , the higher the risk of the 10-year vascular death ( HR 1.37, 95% CI 1.02-1.83; P=0.034). The greater the decrease of INR in patients with ischemic stroke within one year, the higher the 10-year non-vascular death risk ( HR 2.15, 95% CI 1.29-3.59; P=0.004). Conclusions:The 10-year mortality rate of patients with ischemic stroke is higher, and about 3/4 are vascular deaths. The fibrinogen-lowering treatment in the acute stage has no significant effect on the 10-year all-cause mortality of patients with ischemic stroke. The greater the decrease of tPA in half a year, the lower the all-cause mortality; the greater the decrease of D-dimer level at baseline and within 1 year, the higher the 10-year vascular death; the greater the decrease of INR at baseline and within 1 year, the higher the 10-year non-vascular death risk.