Objective To express a mutated insulin gene in HepG-2 cell line to further research of insulin gene therapy. Methods Native human insulin cDNA was obtained from fetus pancreas with RT-PCR. Furin consensus cleavage sequence was introduced into proinsulin cDNA with site-directed mutagenesis (overlap extension PCR), and the new sequence was named as INS/furin. Subsequently, INS/furin was subcloned into the multiple clone sites of plasmid p(G1RE)3BP-1Luc. The new plasmid p(G1RE)3BP-11?furin was identified with the method of enzyme digestion by Hind Ⅲ and EcoR V. HepG-2 cells were transfected with the plasmid p(G1RE)3BP-11?furin by liposome-mediated method. The transfected HepG-2 cells were incubated for 48h in a glucose-containing medium (25mmol/L), and then the conditioned media were collected and HepG-2 cells were harvested respectively. The expression of INS/furin mRNA in transfected HepG-2 cells was examined by RT-PCR, the regained DNA was sequenced and insulin in conditioned media was investigated by radioimmunoassay. Results Two enzymes, Hind Ⅲ and EcoR V, digested p(G1RE)3BP-11?furin, and 2 fragments with length of 260 bp and 4 700bp, were obtained. The 260bp fragment was identified as insulin/furin, indicating that the target gene had been successfully inserted in specific sites. RT-PCR showed that insulin/furin mRNA was expressed in transfected HepG-2 cell, and the regained DNA was confirmed as insulin/furin by sequencing; while insulin was detected by radioimmunoassay in conditioned media. Conclusion The recombinant mammalian expression plasmid p(G1RE)3BP-11?furin has been successfully constructed, and transfected into HepG-2 cells, which therefore may efficiently secrete bioactive insulin.

Objective To investigate different surgical treatments for femoral intertrochateric fractures. To evaluate the results of treatment of intertrochateric fractures with PFNA. Methods 37 patients with femoral intertrochateric fracture were retrospectively reviewed,of which 12 patients were treated operatively with PFNA,25 patients were treated operatively with DHS. Therapeutic efficacy were compared and analyzed. Results All the patients were followed up for 5-18 months(an average of 11 months). All fractures healed in a mean of 3 months. According to Harris'standard of hip-joint fuction,the outcome was classified into excellent,good and poor. In 25 patients with DHS,the good clinical results was 21 cases. In 12 patients with PFNA, excellent and good clinical result was 11 cases. Conclusion PFNAintemal fixation was an effective method for treating the femoral intertrochateric fractures and better than that by DHS.

Objective To investigate the clinical applications of the chain locking-type tension bands. Methods89 cases with patellar bone fractures,27 olecranal fractures patients,18 patients with fractures of surgical neck of humerus,16 patients with dislocation of the acromioclavicular joints and 12 patients with clavicular lateral fractures were treated with chain locking-type tension bands. ResultsTheResultsshowed that all patients wound were postoperative first intention.The healing time of the fractures were 6 ～ 18 months(average 10 months).The Kirschner's pins lapping,steel wire breaks and tension bands out of control were not found. ConclusionCompared with the traditional tension bands,the chain locking-type tension bands had stronger stability,stress distribution more even and less complications.

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.