Objective　To develop a new method to induce the gene transfection in high efficiency for eukaryotic cells in vitro. Methods　 Four kinds of p14ARF gene primarily deleted human carcinoma cell line including H460,A549,U251,and PC-3 were transfected with the human p14ARF expression vector (pCI-neo-p14ARF) by using the new nonliposomal transfection reagent Fugene 6. The efficiency of gene transfer was determined by screening the cells in G418. Results　 After 21 days' selection, G418-resistant clones were shown in all the transfected plate. PCR product of p14ARF gene was positive in all the G418-resistant clones. Cytotoxicity of Fugene 6 was detected. The cell proliferation activity was not affected when it was cultured in a high dose of Fugene 6. Conclusion　 These results demonstrate that Fugene 6 is a rapid, feasible, reproducible, and noncytotoxic gene transfection approach for eukaryotic expression vector in vitro.

Objective　To examine whether wild-type p14ARF gene is a candidate suppressor gene for lung cancer. Methods　Human lung cancer cell lines having various endogenous backgrounds in INK4a, p53 and Rb genes were used as the recipients of the wild-type p14ARF gene. The expression of p14ARF mRNA and protein was detected with RT-PCR, immunohistochemistry and Western blot after G418 selection. Clones which expressed both p14ARF mRNA and protein were identified and selected for further experiements. By comparing with the parental and negative control cells treated with empty vectors, the effects of exogenously transfected p14ARF on cell division rate, cell cycle distribution and morphologic alteration were analyzed. In vivo evaluation of the growth rate was also made with the experiment of nude mice tumor formation. Results　Upon transfection with p14ARF gene, cells were arrested at G1 or G1／G2 phase of cell cycle in three wtp53 lung cancer cell lines and their proliferation rates were also inhibited. Conclusion　Human wild-type p14ARF gene has suppressive effect on abnormal proliferation of lung cancer cells, especially in some wtp53 lung cancer cells, and it might be an ideal candidate for gene therapy of human lung cancer.

Diabetic retinopathy(DR), one of the most common retinal vascular disease, is one of the causes of blindness for people over the age of 50.In the early stage of DR, microvascular cells are damaged, expand, start to leak, form micro hemangioma, then show occlusion, and non-perfusion area come into being, eventually form new blood vessels because of ischemia and hypoxia of retina.Illness develop into proliferative diabetic retinopathy(PDR).With the aggravation of the disease, PDR can cause the formation of fibrovascular membrane, the more serious fibrillation of epiretinal membrane, resulting in traction retinal detachment(tRD).Present studies suggest that aquaporins, the essential component of new blood vessels, including aquaporin 1 and aquaporin 4, play a significant pole in the development of diabetic retinopathy, causing the destruction of blood retinal barrier, inducing retinal edema, even macular edema, and participating in the formation of retinal angiogenesis.

Myopia is the highest incidence of eye disease in society, it threated people`s eye health seriously.At present, many researchers measure the structural changes of retina which caused by the extension of eye axial due to the refractive error according to the optical coherence tomography(OCT).Besides, multifocal electroretinography(mf-ERG) was used to detect the function of retina in myopia.Global flash mf-ERG can evaluate the function of retina including both outer and inner retina.Some researchers focus on the correlation between structural and functional changes in myopia using OCT and mf-ERG.More researches are needed to clarify the structural and functional changes in myopia.

AIM: To retrospectively analyze selection of operation, surgery effect, complications and treatment methods of super high myopia combined with rhegmatogenous retinal detachment( RRD) .
METHODS:A total of 47 cases(47 eyes) with the super high myopia patients combined with RRD who accepted scleral buckling of external pressure, vitrectomy combined silicone oil filling, or intravitreous gas injection were chosen in our hospital from Jan. 1, 2011 to Jan. 1, 2016. Best corrected visual acuity ( BCVA), intraocular pressure, retinal reattached rate, closure rate of holes and the postoperative complications of patients in half year were analyzed.
RESULTS:Half a year postoperation, BCVA of twenty-four cases underwent vitrectomy combined silicone oil filling were improving(71%), and eight cases with scleral buckling surgery were enhanced ( 73%) . Statistical test showed the surgical treatment could significantly improve patients'eyesight, especially PPV with silicone oil, but the BCVA of patients with scleral buckling of external pressure in this study had no obvious significance according to the rank sum test. Compare with other surgery, pars plana vitrectomy( PPV ) combined silicone oil filling had more complications, such as high intraocular pressure and diplopia.
CONCLUSION: The therapy of super high myopia patients with rhegmatogenous retinal detachment need many years' clinical experience, surgical skill of clinician and according to the number, the shape and the site of the retinal holes and area of detached retina, they need to choose the right means of surgery, and timely deal with postoperative complications, in order to improve the patients' postoperative retina, reset rate and visual function.

Objective To understand the disease spectrum of prehospital emergency patients in Beijing, so as to provide basis for rational allocation of prehospital emergency care resources and for improvement of prehospital emergency care. Methods Emergency dispatch database from 120 dispatching command system of Beijing Emergency Medical Center to was used study the disease spectrum of 120 prehospital emergency patients from 2013 to 2017 in Beijing, and were analyzed the differences of disease spectrum among patients of different years, genders and ages, and the time distribution of different diseases. Results From 2013 to 2017 in Beijing, 1 643 375 patients used 120 prehospital emergency care. The top 3 diseases of the disease spectrum were: injury and poisoning (29.63%), diseases of the circulatory system (18.37%), and diseases of the nervous system (15.80%). The sorting of diseases remained unchanged during the 5 years. The disease spectrums were different among different age groups and gender groups. The calls in spring, summer, autumn and winter accounted for 25.23%, 24.97%, 25.05% and 24.75% respectively. Most calls were made from 08 ∶〗00 to 09 ∶〗59 (12.41%), 12 ∶〗00 to 13 ∶〗59 (10.05%), and 14 ∶〗00 to 15 ∶〗59 (9.91%) during a day. The peak months and hours of different diseases were different. Conclusions The demand for prehospital emergency care has been increasing in Beijing in recent years. It is important to strengthen the prevention of injuries among all ages, especially male, and to improve the prevention and response capacity for acute attack of chronic diseases among older people. Prehospital emergency care resources should be rationally allocated according to the peak months and hours of calls.