Objective To evaluate two kinds of practical CT three-dimensional volumetric measurement techniques in assessing atrial septal defects (ASDs) for transcatheter device closure.Methods Retrospective assessment of 50 consecutive patients who underwent ASD closure was conducted.Cardiac CT was performed in them before planned transcatheter ASD closure and postoperative chest radiograph was performed in both posteroanterior and lateral view.Coronary CT angiography was made for the detection of coronary artery disease,and three-dimensional reconstruction of ASD was conducted for determination of the defect size in the GE-workstation.Two kinds of practical CT three-dimensional volumetric measurement techniques,one named CT virtual endoscopy assisted volumetric measurement (CTVE) and another called axial sequence assisted volumetric measurement(CTAS),were used to calculate the major axis and the minor axis of the ASD respectively.Thus,the dimensions of the Equivalent Circle were derived,with circunference and area equal to the elliptic ASD according to the formulation (D =√b4/a2 + a2-b2).The ASDs occluder (ASO) dimension was measured in the lateral chest radiograph,and this postoperative occluder-waist size (POS) value was regarded as the gold standard for the measurement of ASD.The results were compared between CTVE and CTAS,and the correlations were evaluated between them and the gold standard.Results The differences in the major dimensions (-9.05 mm,t =-6.60,P ＜ 0.05),minor dimensions (-4.86 mm,t =-4.39,P ＜ 0.05) and Equivalent circle dimensions (-7.65 mm,t =-6.40,P ＜ 0.05) of ASD between CTAS and CTVE were statistically significant.Though the CTAS cannot provide the en face views of ASDs,the Equivalent Circle dimensions measured by CTAS(22.48 ± 5.59) mm was correlated well with POS (27.07 ± 6.83)mm (Y =1.14X + 1.39,r =0.94,P ＜ 0.01),and a good correlation was found between this Equivalent Circle dimensions and ASO size (Y =1.02X + 6.84,R2 =0.78,r =0.88,P ＜ 0.05).The correlation between the Equivalent Circle dimensions measured by using CTVE (30.13 ± 9.27) mm and POS was poor (Y =0.30X + 17.94,r =0.41,P ＜ 0.01),though it can provide the en face views of ASDs.Conclusion CTVE and CTAS are two complementary techniques of assessing ASDs for transcatheter device closure.

Objective To investigate the efficacy of transcatheter closure of coronary artery fistula (CAF) by transthoracic echocardiography(TTE) and the role of TTE in this therapy. Methods CAF were occluded with transcatheter closure techniques in 17 patients. TTE was performed before and after the treatments. The key points were retrospectively analyzed including: the sites of CAF, the position and diameter of CAF,the shape and position of the devices after the intervention,the residual shunt,and cardiac chamber size. Results Before the therapy,TTE made definite diagnosis of CAF. All patients had lesions in single coronary artery, and the diameter of CAF was 2-14 (6.4 ± 3.5)mm. All patients underwent transcatheter closure successfully. TTE revealed the shunts disappear 1-4 days after the occlusion. During a follow-up period of 1- 29 (11.7± 7.9)months,no residual shunts, no complications, and normal size of thecardiac chambers were recognized by TTE. Conclusions The transcatheter closure of CAF has emerged as a less invasive, safe and effective strategy. Echocardiography has important role in primary screening of patients and the follow-up after the treatments.

Objective To assess the safety and efficacy of balloon dilation of pulmonary valve stenosis with 10 F domestic balloon catheter in children ≥ 10 kg. Methods From May 2009 to June 2014, eighty-three consecutive children with weight ≥ 10 kg and age of (4.5±2.8)(ranged from 1-12) years underwent percutaneous balloon pulmonary valvoloplasty(PBPV) with 10 F domestic balloon catheter. Indication for treatment, procedural details, catheterization data, complication rate, peak-to-peak systolic gradient across the valve and pulmonary insufficiency on echocardiography were respectively analyzed. Forty-four patients were followed up 6-44 months after procedure. Results All procedures were completed successfully. The peak-to-peak systolic gradient across the pulmonary valve decreased from (67.7±26.2) mmHg to (15.4±11.6) mmHg (P < 0.01) immediately after PBPV. Two patients developed reactive infundibular spasm after dilation. They were relieved at 6 months post PBPV. No patient had severe pulmonary insufficiency, tricuspid regurgitation or reintervetion. Conclusions PBPV with 10 F domestic balloon catheter in children with weight≥10 kg is a safe and effective method.

In order to construct the recombinant retrovirus vector of human ngn3 gene and its packaging cell line, we successfully amplified the open reading frame (ORF) of ngn3 gene from human fetal pancreatic tissue by RT-PCR. The PCR products of human ngn3 gene was subcloned into pMD18-T vectors and sequenced. Results showed that its sequence was fully consistent with the ngn3 gene published in GenBank(GenBank Accession No. BC126468). The correct fragment was digested by EcoR I and Hpa I from recombinant pMD18-T vector and inserted into the same restriction enzyme sites of retroviral vector pMSCV-neo. We got recombinant retrovirus vector pMSCV-ngn3, which was identified by double restriction enzyme digestion and then transfected into PT67 cells by lipofectamine 2000. We established the PT67-ngn3 packaging cell line by G418 selection, which was detected by RT-PCR and immunohistochemistry staining. The detection results showed that the Ngn3 expressed at the mRNA and protein level in the packaging cell line. RT-PCR detection and electronic microscope analysis showed that the recombinant retroviral vector pMSCV-ngn3 was packaged into infectious virus particles and released into the supernatant of the cells. These results demonstrated that a PT67-ngn3 packaging cell line was successfully established, and this could facilitate the study of differentiation of the human fetal pancreatic progenitor cells into insulin-producing cells by using the ngn3 gene.
Basic Helix-Loop-Helix Transcription Factors ; biosynthesis ; genetics ; Cell Differentiation ; drug effects ; Cell Line ; Cloning, Molecular ; Fetus ; Genetic Vectors ; genetics ; Humans ; Insulin-Secreting Cells ; cytology ; Molecular Sequence Data ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Open Reading Frames ; genetics ; Pancreas ; cytology ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Retroviridae ; genetics ; metabolism ; Stem Cells ; cytology ; Transfection

In order to construct the recombinant retrovirus vector of bovine sox2 gene and obtain infectious retroviral particles, we successfully amplified the ORF (open reading frame) of bovine sox2 gene from the primodial genital ridges of bovine embryo by RT-PCR. The cDNA of ORF was subcloned to pMD18-T vectors and verified that its sequence was highly homologous to the GenBank counterpart (GenBank Accession No. NM-001105463) by sequencing. The correct fragment was digested by EcoR I/Bgl II from recombinant pMD18-T vector and inserted into the same restriction sites f retroviral vector pMSCVneo. We got recombinant retrovirus vector pMSCV-sox2 which was transfected into PT67 by lipofectamine 2000 with pMIG (including green fluorescence protein) as a control. Flow cytometry analysis showed that its transfected efficiency was 68.3%. Subsequently, we established the stable cell strain by G418 selection which could produce virus. Its viral titer was up to 8.16x10(7) CFU/mL. This greatly facilitates the further study of bovine induced pluripotent stem cells induced from bovine somatic cells by defined factors.
Animals ; Cattle ; Cloning, Molecular ; Genetic Vectors ; genetics ; Mice ; NIH 3T3 Cells ; Open Reading Frames ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Retroviridae ; genetics ; metabolism ; SOXB1 Transcription Factors ; biosynthesis ; genetics ; Stem Cells ; metabolism ; Transfection

AIM:To investigate the effects of adipose-derived stem cells(ADSCs)with overexpression or si-lencing of long noncoding RNA(lncRNA)SNHG8 on the viability,migration,angiogenesis,and the expression of vasoac-tive factors in human umbilical vein endothelial cells(HUVECs).METHODS:Identification of ADSCs derived from morbidly obese patients(O-ADSCs)was conducted using flow cytometry and induction of lipogenesis and osteogenesis.The expression of lncRNA SNHG8 in healthy human ADSCs(H-ADSCs)and O-ADSCs was detected by RT-qPCR.Tran-swell method was used to establish the indirect co-culture system of ADSCs and HUVECs for 48 h,and the cells were di-vided into O-ADSCs+HUVECs group,H-ADSCs+HUVECs group,and HUVECs alone group.The mRNA and protein ex-pression levels of angiotensin Ⅱ(Ang Ⅱ),endothelin-1(ET-1)and endothelial nitric oxide synthase(eNOS)in HUVECs were detected by RT-qPCR and Western blot.The lncRNA SNHG8 overexpression and silencing lentiviruses were con-structed and used to infect O-ADSCs.The indirect co-cultured ADSCs and HUVECs were divided into O-ADSCs-OE-SNHG8+ HUVECs group,O-ADSCs-OE-NC+HUVECs group,O-ADSCs-sh-SNHG8+HUVECs group,and O-ADSCs-sh-NC+HUVECs group.After co-culture for 48 h,the viability,migration and tubule formation of HUVECs were detected by CCK-8,scratch and angiogenesis assays,respectively.The mRNA and protein expression levels of Ang Ⅱ,ET-1 and eNOS in HU-VECs were detected by RT-qPCR and Western blot,respectively.The nitrate reductase method was used to detect the con-tent of NO in HUVECs.RESULTS:(1)The cultured cells were identified as ADSCs.(2)Compared with H-ADSCs,ln-cRNA SNHG8 expression was significantly up-regulated in O-ADSCs(P<0.01).(3)Compared with H-ADSCs+HUVECs group and HUVECs group,the mRNA and protein expression levels of Ang Ⅱ and ET-1 in HUVECs in O-ADSCs+HU-VECs group were up-regulated(P<0.01).(4)Overexpression of lncRNA SNHG8 in O-ADSCs enhanced the viability,mi-gration and tube formation ability of HUVECs,up-regulated the mRNA and protein expression levels of Ang Ⅱ and ET-1,down-regulated the mRNA and protein expression levels of eNOS,and decreased the content of NO in HUVECs(P<0.05).However,silencing of lncRNA SNHG8 in O-ADSCs exerted opposite results(P<0.05).CONCLUSION:(1)The O-ADSCs can promote endothelial cell viability,migration and tubule formation through paracrine effects.(2)The O-ADSCs with overexpression of lncRNA SNHG8 promote the imbalance of diastolic and contractile factors secreted by endo-thelial cells,and induce the dysfunction of vascular endothelial cells.