Objective: To investigate the inhibitory effects of RNA silencing via adenovirus mediated vascular endothelial growth factor(VEGF) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo. Methods: PCR method was used to construct a pAd Easy/VEGF adenovirus vector containing enhanced green fluorescent protein (EGFP) gene and expressing VEGF shRNA. The 293 cells were transfected with the linearized pAd Easy/VEGF using Lipofectamine2000. Then lung adenocarcinoma cells A459 were transfected with the constructed vector. The EGFP expression was detected by fluorescent microscopy and flow cytometry. VEGF mRNA expression was examined by RT-PCR and Western blotting. The cell growth was observed with MTT method and the growth curve was plotted. Meanwhile, nude mice were transplanted with A549 cells to establish tumor models and the growth of tumors were observed. Results: The recombinant pAd-Easy carrying shRNA targeting VEGF had been constructed and the aim sequence had been obtained. The transfection efficiencies in pAd-Easy/VEGF and blank vector transfected A549 cells were 100% and 99.7%, respectively. RT PCR and Western blotting showed a remarkable decrease of VEGF expression in the pAd-Easy/VEGF group compared with normal saline group. The tumor growth in pAd-Easy/VEGF group was obviously slowed down and the weight and volume of tumors were both significantly lower than those of the control group (all P<0.01). Conclusion: The shRNA targeting VEGF constructed in the present study can efficiently decrease the VEGF expression in A549 cells in vitro and suppress the growth of A549 cells in vivo.

Objective: To evaluate the protective effects of adenovirus-mediated gene transfer of soluble complement receptor type 1 (sCR1) on acute myocardium ischemia in mice. Methods: Twenty-seven SD rats were subjected to left anterior descending coronary artery (LAD) occlusion (30 min) and reperfusion. In the treatment group (n = 14), a mixture of adenovirus carrying sCR1 and LacZ was injected into the ischemic zone(100 μl, 1010 pfu) 5 min before reperfusion; in the control group, only adenovirus carrying LacZ was injected (n = 13). Echocardiography was performed 2 weeks later, followed immediately by pathologic examination. Results: Echocardiographic results of the treatment group were better than those of the control group (P<0.05) 2 weeks after the operation, especially when it comes to the size of infarction area (P<0.01). H-E staining showed more surviving myocardial cells in the treatment group than in the control group. Conclusion: Transfer of sCR1 gene by adenovirus can reduce the size of infarction area and preserve heart function in a long-term period.

Objective · To investigate the prevalence of passive smoking among pregnant women in Shanghai and to analyze its influencing factors and to provide the basis and suggestion for making related intervention measures to reduce the harm of secondhand smoke on the health of pregnant women. Methods · From July to October 2014, pregnant women and their spouses were recruited for a questionnaire investigation at the antenatal care clinics of 3 maternity hospitals and 3 community hospitals in Shanghai. The questionnaire included demographic information, family basic situation and cognitive status of passive smoking. Results · The passive smoking rate before pregnancy in the 2831 pregnant women, who were involved in the investigation, was 17.1%. The passive rate during pregnancy was 7.8%. The difference between these two rates was statistically significant (P<0.05). 71.0% of the pregnant women have passive smoking for 15~59 min each day. Relative to home and work place, the passive smoking rate of pregnant women in public place was the highest (P<0.05). The passive smoking rate of pregnant women was significantly associated with the educational level of smoking spouses and the attitude of pregnant women when they faced the smokers. Conclusion · The passive smoking rate of pregnant women is influenced by the educational level of their spouses and the pregnant women's feedback to the smokers face to face. Public places are the most important exposure places to secondhand smoke for pregnant women. In order to reduce the harm from passive smoking to pregnant women and their fetuses, the related measures of smoking control at home education and smoking ban in public place should be made.

Objective To study the expression and significance of miR-26a in intrahepatic cholangiocarcinoma.Methods The expression of miR-26a in 46 intrahepatic cholangiocarcinoma(ICC) tissues and peritumoral tissues was detected by quantitative real time polymerase chain reaction (qRT PCR).The intrahepatic eholangiocarcinoma cell line HCCC-9810 and RBE were transfected with miR 26a mimics and miR 26a inhibitors,respectively,by lipofectamine 2000.The growth curves were constructed by the CCK 8 method.The migration and invasion ability was demonstrated by wound healing and transwell assay.The cell cycle was analyzed by flow cytometry.The potential mechanism was illustrated by Western blotting.Results For the 46 ICC tissues and peritumoral tissues,miR 26a levels were significantly higher in the tumor tissues than in the peritumoral tissues (P＜0.05).Vascular invasion,TNM Ⅲ～Ⅳ stage and lymph node metastasis were significantly associated with high miR 26a expression levels (P＜0.05),but gender,age,tumor amounts,tumor encapsulation,tumor diameter and tumor differentiation showed no significant association (P＞0.05).Enhanced cell proliferation,migration and invasion ability,accelerated G0/G1 phase to S phase transition,activated AKT by PTEN suppression were observed in HCCC-9810 cells with up regulation of miR-26a.Conversely,cell proliferation,migration and invasion ability was inhibited,G0/G1 phase was blocked and AKT was restrained by PTEN increase wkh down regulation of miR-26a in RBE cells.PTEN mRNA in versely correlated with the miR-26a level (r=-0.8272,P＜0.01).Patients with a high miR-26a expression had significantly poorer overall survival (P＜0.05).A high miR 26a exprcssion,multiple tumors and lymph node metastasis were independent prognostic factors of overall survival (P＜0.01).Conclusion Overexpression of miR-26a in intrahepatic cholangiocarcinoma correlated with clinicopath ological features and overall survival,and it potentially promoted tumor proliferation and metastasis through the PTEN/AKT pathway.

Child ; Chondrosarcoma, Mesenchymal ; diagnosis ; drug therapy ; pathology ; Diagnosis, Differential ; Humans ; Male ; Pelvic Neoplasms ; diagnosis ; drug therapy ; pathology ; Tomography, X-Ray Computed ; Treatment Outcome

Recently,study on cencer stem cells has been a focus of study.Cancer stem cell is a small population of cencer cells possessing the properties of stem cells:self-renewal,differentiation and proliferation.To date,the existence of cancer stem cells has been proven in acute and chronic myeloid leukemia,breast cancer,brain tumors,liver cancer and colon cancer,etc..In this article we reviews the current progress on cancer stem cells,including the defination,existing evidence,research methods, and challenges in clinical application.

AIM:To investigate the effect of intravitreal injection of anti-vascular endothelial growth factor (VEGF) drugs combined with other methods in the treatment of Coats disease.METHODS:Selected 13 patients 13 eyes with Coats disease in our hospital from May 2013 to May 2016.All eyes were treated with intravitreal injection of ranibizumab and combined with scleral drainage, laser photocoagulation and so on.We observed visual acuity and retinal reattachment.RESULTS:In 13 eyes, the treatment of 4 eyes with intravitreal injection of ranibizumab and combined with scleral drainage, 3 eyes combined laser photocoagulation, 6 eyes combined vitrectomy, membrane peeling, freezing, silicone oil filling or other treatments.Eyeball retention rate was 100%.Visual acuity at 6mo after treatment significantly improved compared with before treatment (P<0.05).The visual acuity was ≥0.05-0.1, 0.01-<0.05, counting finger, light perception and hand moving before treatment in 0, 0, 38%, 38% and 23%, after treatment were 23%, 23%, 38%, 15% and 0.Complete retinal reattachment was achieved in 5 eyes, accounting for 38%;basic reset in 3 eyes, accounting for 23%;5 eyes were not reset, accounting for 38%.CONCLUSION:Intravitreal injection of anti-VEGF drugs combined with other methods for the treatment of Coats disease have a certain effect, can improve or maintain the limited visual function, avoid enucleation of eyeball due to disease progression.

Trachoma, a contagious keratoconjunctivitis ( KC ) , caused by Chlamydia trachomatis infection, is rife in 57 countries in the world at present. The World Health Organization ( WHO) listed the global alliance to eliminate blinding trachoma by 2020 as one of top priorities of its blindness prevention in 1998. A simplified classification system for identifying and naming trachoma, designated by WHO, and the SAFE strategy based on community intervention were extended continuously in the world in 10 years since then. The trachoma prevalence trend has showed a change compared with that in the past. China has launched the blindness prevention action, aimed to eliminate blinding trachoma by 2016. In this paper, we reviews progress in diagnosis, treatment and epidemic of trachoma since the extension of the SAFE strategy.

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Objective To investigate the immunological activity change of regulatory T cells (Treg) and discuss its significance in the outcomes of patients with multiple organ dysfunction syndrome (MODS) and severe burn. Methods A total of 106 patients with total burn surface area (TBSA) larger than 30% were included in the study and randomly divided into three groups according to the burn area: Group Ⅰ (TBSA of 30%-49%, n = 41), Group Ⅱ (TBSA of 50% -69%, n = 34) and Group Ⅲ (TBSA of 70%-99%, n = 31). According to the development of MODS, patients were divided into MODS group (n =21) and non-MODS group (n =85). The patients with MODS were further divided into non-survival group (n = 16) and survival group (n = 5) based on their outcomes. Healthy volunteers were served as normal control (n = 25). Peripheral blood samples were collected at days 1,3,7, 14 and 21 postburn. The immunomagnetic separation technique was applied to separate and purify CD4+ CD25+Tregs in peripheral blood, and phenotypes (CTLA-4) were analyzed by flow cytometry and the contents of interleukin-10 released in the supernatants were determined by ELISA. Results Expression of CTLA-4 and level of IL-10 were significantly increased in burn patients compared with normal control group, with statistical differences. The expression of CTLA-4 and level of IL-10 were significantly increased in patients with severe burns at all time points. The expression of CTLA-4 and level of IL-10 in MODS group were much higher than those in non-MODS group at days 3-21 postburn (P ＜ 0.01). Among the MODS patients, the expression of CTLA-4 and level of IL-10 in the survival group were obviously lower than those in the non-survival group at days 3-21 postburn (P ＜ 0.05 or P ＜ 0.01). Conclusions After severe burn injury, expressions of the markers on CD4 + CD25 + Treg surface and secretion of cytokines produced by CD4 + CD25 + Tregs show significant difference in patients with different born areas, MODS development and survival state. CD4 + CD25 + Treg may play an important role in the pathogenesis of immunoregulation, MODS and mortality of burn patients through secretion of inhibitory cytokines.