Objective RNA interference refers to post-transcriptional gene silencing caused by double strands RNA.To investigate the effect of EGFR receptor on esophageal carcinoma,the expression vector of HER4 gene targeted small interfering RNA was constructed to observe its silencing effect in human esophageal carcinoma cell line Eca-109,in order to find a promising method for the gene therapy of this disease.Methods Two complementary oligo DNA strands targeting HER4 gene were designed and synthesized according to the principles of designing siRNA.After annealing,oligo DNAs were inserted into SUPER.neo+gfp vector,then enzyme digestion analysis and DNA sequencing were applied.After transfecting it into human esophageal carcinoma cell line,we detected the level of expression of HER4 gene through real-time quantitative PCR and Western Blot.Results The enzyme digestion analysis and DNA sequencing show that HER4 gene targeted small interfering RNA and its expression vector were constructed successfully,and after transfection,the expression of HER4 gene in esophageal carcinoma cell line was suppressed greatly.Conclusion HER4 gene targeted small interfering RNA and its expression vector were constructed successfully,and could decrease the expression of HER4 gene in Eca-109 cell line,which laid the foundation for the following experiment.

The voltage-gated potassium channels Kvl.5 is widely expressed in the plasma membranes of numerous tumor cells and it can contribute to a variety of cellular functions such as proliferation,ap optosis,and cell cycle.Several types of antineoplastic drugs can change the expression of Kv1.5 and then affect the biological processes.Kv1.5 is identified as a novel target for therapy in human cancer.The researches of Kv1.5 will contribute to gain a further understanding of the molecular mechanism of tumor and provide therapeutic opportunity for the prevention and treatment of cancer.

The ultrasonographic appearances of osteosarcomas and the roles of ultrasonography in the diagnosis and surgical staging of osteosarcomas were investigated. A comparative study was performed on 45 cases of osteosarcomas by ultrasonography and radiography. Bony changes, periosteal reaction and soft tissue mass were evaluated for each lesion. The results showed that ultrasonography revealed a solid mass around bone in 42 patients, bone destruction in 24 patients and periosteal reaction in 16 patients. Plain radiographs showed bony changes in 44 patients and no bony change in remaining one patient, shadowing of soft tissue swelling in 30 patients, and pulmonary metastases in 3 patients. Surgical biopsy and pathological examination confirmed osteosarcoma in all 45 patients. Soft tissue mass was confirmed in 42 patients surgically. The diagnostic accuracy of soft tissue masses by ultrasonography and radiography was 100 % (42/42) and 71.4 % (30/42), respectively. The positive rate of ultrasonography and radiography in displaying bony changes was 53.3 % (24/45) and 97.8 % (44/45), respectively. In conclusion, in the detection of soft tissue mass of osteosarcoma, ultrasonography is superior to radiography, and in displaying bony changes of osteosarcomas, radiography is superior to ultrasonography. So it may come to a conclusion that plain radiography combined with ultrasonography can completely display the bony and soft tissue lesion of osteosarcomas.

Objective To explore the associations of depressive symptoms with functional status and frailty in elderly outpatients.Methods A total of 297 geriatric outpatients(aged 65 years and over) from Zhejiang Hospital were recruited in the cross-sectional study from January 2014 to December 2015.We gathered general information,and evaluated depressive symptoms by Geriatric Depression Scale-15 (GDS-15),cognitive function by mini-mental state examination(M MSE),frailty by clinical frailty scale(CFS),activities of daily living (ADL)by Barthel index,instrumental activities of daily living (IADL),balance,POMA and gait by Tinetti-performance oriented mobility assessment (Tinetti-POMA),grip strength and 4m gait speed by 4-meter walk gait speed test.According to the GDS-15 scores,297 geriatric outpatients were divided into a depression symptom group(n=35,GDS-15 ≥6)and a nomdepressive symptom group(n=262,GDS-15＜6).The frailty and functional status were compared between two groups by SPSS 23.0.Results As compared with non-depressive symptoms,the depressive symptoms group had higher clinical frailty scale(CFS),lower body mass index(BMI),lower cognitive function and poorer grip strength and balance(all P＜0.05).There were significantly negative correlations of Geriatric Depression Scale-15 (GDS-15) with ADL,IADL,gait,balance,POMA and grip strength(r=-0.165、-0.154、-0.216、-0.291、-0.305、-0.314,All P＜ 0.05),while there were significantly positive correlations with CFS score,gait speed(r=0.256、0.198,both P＜0.05).The more severe the frailty was,the higher the risk of depressive symptoms was(OR=3.650,95％ CI 1.611-8.271).Conclusions The cognitive and physical functions in the elderly with depression symptoms are poorer as compared with the elderly without depression symptoms.Elderly outpatients with more severe frailty have a higher risk for depressive symptoms.

A 53-year-old woman presented with arthralgia and dyspepsia on exertion.Symmetrical joint swelling and pain of bilateral hands with pigmentation sedimentation in the articular surfaces,dry cough and short of breath were the main clinical manifestations.Elevated levels of erythrocyte sedimentation rate (89 mm/1 h),C-reactive protein (64.45 mg/L),IgG (31.22 g/L) and IgA (5.44 g/L).Rheumatoid factor was positive and anti-cyclic citrullinated peptide antibody was negative.Chest high-resolution computed tomography scan found that diffusely distributed lung nodules,round cysts with varying sizes and patchy ground glass opacities.A significant plasma cell infiltration and amyloid deposition were seen in lung tissue.The patient was finally diagnosed as rheumatoid arthritis,interstitial lung disease and pulmonary nodules.Combination therapy of prednisone 7.5 mg qodpo,thalidomide 50 mg qdpo,tocilizumab 560 mg iv once per month for 1 month.Chest computed tomography showed stable lung nodules,however,pulmonary interstitial lesions gradually aggravated.

Objective To detect the expression of HERG (human ether-à-go-go-related gene ) potassium channel in human osteosarcoma,and explore the effects of silencing HERG by small interfering RNA (siRNA)on the proliferation and apoptosis of osteosarcoma cells and the mechanisms responsible for HERG regulation.Methods The expressions of HERG in osteosarcoma MG-63 cells and tissues were detected by reverse transcription polymerase chain reaction (RT-PCR),Western blotting and immunohistochemistry.Next, osteosarcoma cells were divided into three groups:HERG-siRNA group,control-siRNA group and blank group. CCK-8,colony formation,flow cytometry and Tunel assay were used to measure the proliferation and apoptosis of the osteosarcoma cells.Finally,Western blotting analysis was performed to detect the expression of nuclear factor-κB (NF-κB)pathway in osteosarcoma cells treated with HERG siRNA.Results Osteosarcoma cells and tissues were found to highly express HERG.Inhibition of HERG in the osteosarcoma cells significantly inhibited the cell proliferation and induced cell apoptosis.Compared to control-siRNA group or blank group,HERG-siRNA could inhibit the proliferation of MG-63 cells significantly [HERG-siRNA group:(75.34 ± 4.45)%;compared to control-siRNA group:(100.60 ±5.31)%;t =3.64,P =0.007;compared to blank group:(100.00 ±5.66)%;t =3.43,P =0.009].The similar results were obtained from colony formation assay (HERG-siRNA group:134.30 ±11.82;compared to control-siRNA group:225.30 ±11.56;t =5.51, P =0.002;compared to blank group:232.80 ±12.21;t =5.80,P =0.001).HERG-siRNA transfected MG-63 cells demonstrated a significant increase of apoptotic rate compared to control-siRNA transfected cells or untreated cells [HERG-siRNA group:(28.10 ±2.21 )%;compared to control-siRNA group:(9.36 ± 2.42)%;t =5.72,P =0.005;compared to blank group:(10.92 ±2.51)%;t =5.14,P =0.007].This resultwas further confirmed by Tunel assay.The cells transfected with HERG-siRNA (31.57 ±2.08)% dem-onstrated extensive apoptosis,compared with the control-siRNA group [(10.35 ±1.82)%;t =7.69,P =0.002)]or blank group [(7.96 ±0.88)%;t =10.48,P =0.001].Silencing HERG gene down-regulated the cIAP-1,XIAP,Bcl-2,Survivin,P-IκBαand NF-κB p65 expression,compared to the control groups. Conclusion HERG is highly expressed in osteosarcoma.HERG silencing can suppress osteosarcoma progres-sion through NF-κB pathway and suggest that HERG may be a novel molecular target for osteosarcoma therapy and diagnosis.

Objective:To investigate the expression of Col9a1 gene in ankle tissue of rat model of idiopathic congenital talipes equinovarus (ICTEV) . Methods:ICTEV rat model was established. Immunofluorescence staining and immunohistochemical dyeing technology were used to detect the expression of Col9a1 protein in ankle tissue of model group and control group. Results:The expression level of Col9a1 protein in ankle tissue in model group was higher than that in the control group (P<0.05),Col9a1 gene expressed primarily in soft tissue and cartilage membrane. Conclusion:The increasing expression level of Col9a1 gene in ankle tissue of ICTEV rats may be related to clubfoot deformity.

Abstract Teen E sports and their negative impact should not be underestimated. The effects of gaming, as an emerging sport, should be examined to determine whether it plays an active role in contributing to the onset of adolescent depression. Therefore, this study outlines the development status of E sports, analyzes the influence of E sports on adolescent depression, and highlights the hidden concerns that E sports pose for the mental health development of adolescents. While taking into account the advantages of E sports and exercise intervention programs, an emphasis should be placed on the prevention of depression, and feasible suggestions and countermeasures should be put forward to promote E sports, while at the same time safeguarding the healthy and orderly development of adolescent mental health.

The ultrasonographic appearances of osteosarcomas and the roles of ultrasonography in the diagnosis and surgical staging of osteosarcomas were investigated. A comparative study was performed on 45 cases of osteosarcomas by ultrasonography and radiography. Bony changes, periosteal reaction and soft tissue mass were evaluated for each lesion. The results showed that ultrasonography revealed a solid mass around bone in 42 patients, bone destruction in 24 patients and periosteal reaction in 16 patients. Plain radiographs showed bony changes in 44 patients and no bony change in remaining one patient, shadowing of soft tissue swelling in 30 patients, and pulmonary metastases in 3patients. Surgical biopsy and pathological examination confirmed osteosarcoma in all 45 patients.Soft tissue mass was confirmed in 42 patients surgically. The diagnostic accuracy of soft tissue masses by ultrasonography and radiography was 100 % (42/42) and 71.4 % (30/42), respectively. The positive rate of ultrasonography and radiography in displaying bony changes was 53.3 % (24/45) and 97.8 % (44/45), respectively. In conclusion, in the detection of soft tissue mass of osteosarcoma, ultrasonography is superior to radiography, and in displaying bony changes of osteosarcomas, radiography is superior to ultrasonography. So it may come to a conclusion that plain radiography combined with ultrasonography can completely display the bony and soft tissue lesion of osteosarcomas.

Objective To explore the expression of ether à go-go 1 ( Eag1) in human osteosarcoma and its molecular mechanisms of regulating the malignant phenotype of osteosarcoma .Methods The expression levels of Eag 1 in osteosarcoma cell lines and human osteosarcoma tissues were detected by reverse transcription polymerase chain reaction ( RT-PCR), western blot analysis and immunohistochemistry .The small interfering RNA ( siRNA) was used to inhibit the expression of Eag 1.The abilities of proliferation and invasion in osteosarcoma cells transfected with Eag 1 siRNA were determined by CCK-8, colony formation assay, transwell assay and wound healing assay .The osteosarcoma xenograft model of nude mouse was established and tumor growth curve was drawn .Western blot analysis was performed to detect the expression of vascular endothelial growth factor ( VEGF) and signal transducer and activator of transcription 3 ( STAT3) in osteosarcoma cells transfected with Eag 1 siRNAs.Results Eag1 was overexpressed in the osteosarcoma cells and tissues .Compared with the scrambled siRNA group , the cell survival rates of Eag 1 siRNA1 and Eag1 siRNA2 groups of the two cell lines were significantly lower .[ MG-63 cells: scrambled siRNA group (100.0±4.65)%, Eag1 siRNA1 group (63.57±3.89)%, and Eag1 siRNA2 group (54.13±3.70)%;Saos2-cells:scrambled siRNA group (100.00±5.46)%, Eag1 siRNA1 group (56.70±5.34)%, and Eag1 siRNA2 group (40.27±5.28)%(P<0.001 for all)].Similar results were obtained from colony formation assay .The colony formation rates of MG-63 cells: the scrambled siRNA group was (92.00±3.46)%, Eag1 siRNA1 group (60.00±3.06)%, and Eag1 siRNA2 group (53.67±2.40)%; the colony formation rates of Saos-2 cells: the scrambled siRNA group was (92.00±5.57)%, Eag1 siRNA1 group (52.33±5.13)%, and Eag1 siRNA2 group (41.67±27.3 )%.Compared with the scrambled siRNA group , P<0.001 for all.The tumor volumes of osteosarcoma xenograft in the Eag 1 siRNA1 and Eag1 siRNA2 groups were significantly smaller than that in the scrambled siRNA group after 10 days treatment (P<0.01 for all).The invasion assay data showed that MG-63 and Saos-2 cells transfected with Eag 1siRNAs exhibited the ability of cell invasion , when compared with the cells transfected with scrambled siRNA .( Invasive cell number of MG-63 cells:the scrambled siRNA group was 134.00 ±3.61, Eag1 siRNA1 group 105.20 ±2.52, and Eag1 siRNA2 group 91.00±3.01; Invasive cell number of Saos-2 cells: the scrambled siRNA group was 132.30±3.23, Eag1 siRNA1 group 114.30±3.48, and Eag1 siRNA2 group 82.67±6.33.Compared with the scrambled siRNA group, P<0.01 for all.The migration rates were (62.48±1.83)%, (35.98±1.23)%and (32.30±1.20)%in the three groups of MG-63 cells, and (70.15±1.42)%, (41.38±1.34)%and (32.40±1.92)%in the three groups of Saos-2 cells, respectively.Compared with the scrambled siRNA group , P<0.001 for all.Notably, the expression levels of VEGF decreased evidently after Eag 1 siRNAs transfection , paralleled with reduction in the expression levels of STAT 3.Conclusions Eag1 may promote osteosarcoma cell proliferation and invasion by targeting STAT 3-VEGF pathway and may be a potential therapeutic target for osteosarcoma .