Objective To construct cDNA library of the treated Changliver cell by switching mechanism at 5′ end of RNA transcript (SMART) technique and analyze its quality.Methods cDNA of Changliver cell was aquired with reverse transcription polymerase chain reaction (RT-PCR) and long-distance PCR (LD-PCR),then the cDNA library was constructed with SMART cDNA library construction kit.Results Through testing,the high quality cDNA library containing full length cDNA of Changliver cell had been constructed.The titer of the amplified cDNA library was 4.5 × 1010 pfu*ml-1 and the average exogenous inserts of the recombinants was 1.5 kb.Conclusion These results suggest that the Changliver cell cDNA library has a high quality and lays a solid foundation for researching on Changliver cell and screening

Objective:To explore the role of N-cadherin and β-catenin in the formation of spinal commissural axon projection during chicken embryonic development.Methods:Fertilized eggs were cultured for three days(stage22),N-cadherin orβ-catenin inter-ference plasmid was injected into the neural tube and in vivo electroporation was performed.Three days after the electroporation, embryos were collected,fixed with 4%PFA,embeded with OCT,and cut into frozen sections.Four groups ( knockdown of N-cadherin orβ-catenin or both of them,and control) were included in this study.Immunohistochemistry method was used to analyze the protein expression result of N-cadherin or β-catenin.The changes of spinal commissural projections were observed with GFP fluorescence.Results:During chicken embryonic development,knockdown of N-cadherin inhibited the expression of β-catenin in the spinal cord.The commissural nerve fibers projecting to the contralateral side of the spinal cord was impaired after knockdown of N-cadherin or β-catenin;this phenotype was similar after knocking down both of them.Conclusion: N-cadherin is implicated in the formation of spinal commissural projection in the developing spinal cord,possibly viaβ-catenin.

ObjectiveTo measure the perfusion parameters of liver fibrosis of dog model with 64-slice spiral CT,and compare with positive control to reflect the degree of liver fibrosis,and analyze the correlation with VEGF values.Methods Liver fibrosis was induced in dogs by intraperitoneal injection of CC14 and high fat diet.CT perfusion scan and liver biopsies were performed.The perfusion parameters were measured according to the liver fibrosis models,and the correlation with VEGF values was analyzed.ResultsThe animals in experimental group were successfully induced different degree of liver fibrosis.In normal group,the hepatic artery perfusion,portal vein perfusion and total hepatic blood flow were (28.25 ±2.19)ml/(min · 100 g),(53.53 ± 10.71)ml/(min · 100 g) and (81.78 ± 18.56) ml/(min · 100 g).The PVP and TLP values of the liver fibrosis models of 0 - 4 stages declined gradually,and it had statistical significance,while the HAP values increased.The PVP and TLP values were positive correlated with fibrosis stage.The VEGF values of the pathological models of 0 - 4 stages ascended significantly.Conclusions The dog liver fibrosis models that are similar to human hepatic fibrosis have been successfully induced by low dose CC14 and high fat diet.CT perfusion can be used to monitor the tendency of the hemodynamic in different degree of fibrosis.VEGF may play an important role during liver fibrogenesis.

Objective The objective of this study was to utilize proton magnetic resonance spectroscopy (1H-MRS) to assess metabolites in cerebellar nuclei in unmedicated patients with insomnia disorder. Methods 1H-MRS was performed on cerebellar nuclei in 23 unmedicated patients with insomnia disorder (insomnia group) and 18 normal sleepers (control group). N-acetylaspartate (NAA), choline-containing compound (Cho) and creatine (Cr) were measured and the ratios of NAA/Cr and Cho/Cr were determined.Pittsburgh Sleep Quality Index (PSQI) and Insomnia Severity Index (ISI) were used to assess the subjective sleep quality and insomnia severity of all subjects, while State-Trait Anxiety Inventory (STAI) and Beck Depression Inventory (BDI) were used to assess the levels of anxiety and depression of all subjects. Sleep parameters of all subjects were measured by polysomnography (PSG). Results Mean NAA/Cr ratio of right cerebellar nuclei in insomnia group was significantly lower than that in control group (1.72±0.37 vs. 2.03±0.50, t=2.280, P=0.028). Mean NAA/Cr ratio of right cerebellar nuclei was significantly higher than that of left cerebellar nuclei within control group (2.03±0.50 vs. 1.68±0.21, t=3.386, P=0.004). There was no significant difference with regard to NAA/Cr ratio between bilateral cerebellar nuclei within insomnia group (t=1.416, P=0.171). Across all subjects, PSQI global scores (r=-0.369, P=0.018), and sleep latency (r=-0.437, P=0.004) and number of awakenings after sleep onset (r=-0.432, P=0.005) measured by PSG were negatively correlated with NAA/Cr ratios of right cerebellar nuclei, while percentages of stage 3 sleep (r=0.377,P=0.015) measured by PSG were positively correlated with NAA/Cr ratios of right cerebellar nuclei,respectively. Conclusion Patients with insomnia disorder have a hemispherically lateralized metabolic disturbance of NAA/Cr in right cerebellar nuclei,indicating that patients with insomnia disorder have neuronal damage in right cerebellar nuclei.

Objective: This study’s objective is to assess the efficacy and safety of Pulsed Magnetic Therapy System (PMTS) in improving insomnia disorder. Methods: Participants with insomnia disorder were randomly assigned to receive either PMTS or sham treatment for four weeks (n= 153; PMTS: 76, sham: 77). Primary outcomes are the Insomnia Severity Index (ISI) scores at week 0 (baseline), 1, 2, 3, 4 (treatment), and 5 (follow-up). Secondary outcomes are the Pittsburgh Sleep Quality Index at baseline and week 4, and weekly sleep diary-derived values for sleep latency, sleep efficiency, real sleep time, waking after sleep onset, and sleep duration. Results: The ISI scores of the PMTS group and the sham group were 7.13±0.50, 11.07±0.51 at week 4, respectively. There was a significant group×time interaction for ISI (F3.214, 485.271=24.25, p<0.001, ηp 2=0.138). Only the PMTS group experienced continuous improvement throughout the study; in contrast, the sham group only experienced a modest improvement after the first week of therapy. At the end of the treatment and one week after it, the response of the PMTS group were 69.7% (95% confidence interval [CI]: 58.6%–79.0%), 75.0% (95% CI: 64.1%–83.4%), respectively, which were higher than the response of the sham group (p<0.001). For each of the secondary outcomes, similar group×time interactions were discovered. The effects of the treatment persisted for at least a week. Conclusion PMTS is safe and effective in improving insomnia disorders.