Objective:To explore the angiographic distribution of the anomalous ostium of right coronary artery (RCA) and improve the technical tips for catheter incubation during right coronary angiography.Methods:A total of 24 861 patients who underwent coronary angiography in Zhongshan Hospital,Fudan University from May 2015 to May 2017 were reviewed,and the cases with anomalous RCA origin were identified.The aortic root at left anterior oblique 40°-45° view during coronary angiography was set as an imaginary plane,which was divided into four quadrants.The ostia distributions of anomalous RCAs,the image features and the technical essentials were analyzed.Results:Totally 147 cases had anomalous RCA origin and all underwent coronary angiography successfully.Among them,67 patients (45.6％,Type A) had an ectopic RCA origin from left sinus of valsalva (LSOV),48 patients (32.7％,Type B) had an anomalous RCA origin from left side of ascending aorta,and 32 patients (21.7％,Type C) had an anomalous RCA origin from right side of ascending aorta.In angiography image,some special signs named as "sea gull sign" and "horn sign" were observed in cases with anomalous RCA origin from LSOV or aortic sinus crest.Such as transposition of the heart and widened aortic root,the anomalous RCA originated from ascending aorta near coronary sinus in angiography image were actually originated from LSOV in some cases.Conclusions:The anomalous RCA arising from LSOV is the most common variation,followed by arising from ascending aorta.The RCA arising from ascending aorta above right coronary sinus is relatively rare.Understanding of the distribution regularity and image features of ectopic RCAs is helpful for coronary angiography and intervention.

OBJECTIVETo investigate the optimal starvation conditions of human umbilical vein endothelial cells (HUVECs) and establish a highly efficient and stable method for separating HUVECs.

METHODSHUVECs harvested from human umbilical cords by digestion with 0.1% collagenase II for 15 min were cultured in endothelial culture medium (ECM) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factor (ECGS) and 1% penicillin/streptomycin solution(P/S) at 37 degrees celsius; in 5% CO2. The cells were observed for cell morphology under an inverted microscope and identified with immunofluorescence assay. The purity of HUVECs was detected using flow cytometry (FCM). The cell cycles of HUVECs cultured in the presence of 0, 0.1%, 0.5%, and 1% FBS for 0, 6, 12, 18, and 24 h were analyzed with flow cytometry.

RESULTSs The purity of HUVECs harvested by digestion with 0.1% collagenase II reached 99.67%. The primary HUVECs showed a cobblestone or volute appearance in vitro. Immunocytochemistry showed that HUVECs highly expressed VIII-related antigen. Cell culture in the presence of different concentrations of FBS for 6 h resulted in 70% G0/G1 phase cells, which increased to 80%-90% at 12 h of cell culture, and further to around 95% at 18 and 24 h.

CONCLUSIONDigestion with 0.1% collagenase II can obtain high-purity primary HUVECs. Culturing HUVECs in serum-free medium for 12 h can result in a high purity (over 80%) of G0/G1 phase cells.

Cell Culture Techniques ; Cell Cycle ; Cells, Cultured ; Culture Media ; chemistry ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Matrix Metalloproteinase 8 ; chemistry ; Serum

Objective To investigate the expression of vasoactive intestinal peptide (VIP)in gastric adenocarcinoma,and to evaluate the correlation of VIP level with clinical pathologic parameters. Methods The level of VIP in sera from gastric adenocarcinoma patients and healthy people was investigated by ELISA. Moreover,the differential gene expression between gastric adenocarcinoma,gastric dysplasia,and the corresponding normal gastric mucosa were determined by RT-PCR. Western Blot was also used to measure the expression of VIP in the gastric odenouarcinoma and the normal gastric mucosa. Results The serum level of VIP was (5.794±0.014) ng/ ml in normal control and was (14.437±0.825) ng/ml in gastric adenocareinoma patients,showing significant difference (P<0.05). Meanwhile,the V/B of gastric adenocarcinoma tissues was greater than that of gastric dysplasia and the corresponding normal gastric mucosa (P < 0.01 ),the values of V/B were 1.5261±0.3028,0.9334±0.2872,and 0.9051±0.2794,respectively. The values of V/B between normal gastric mncesa and gastric dysplasia were not different significantly (P0.05).There were significantly negative correlation between the VIP mRNA expression of the differentiation degree of tumor( P < 0.05). The VIP mRNA expression was higher in gastric adenocarcinoma with lymph node metastasis than that without lymph node matsstsis (P<0.05).The VIP protein expression of the gastric adenouarcinoma tissues was greater than that of normal control. Conclusion This findings provide a direct evidence to support the possibility that VIP play a cofactor role in the pathogenesis of gastric adenccareinoma.

OBJECTIVETo investigate the expression of CD160 on the surface of human natural killer (NK) cells and its possible relationship with hematological malignancies.

METHODSCD160 expression on human leukemia cell line NK92 cells was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The proliferation characteristics and cell surface markers of this cell line were determined. Cytotoxicity of NK92 against 2 human myeloid leukemia cell lines, K562 and THP-1 was analyzed ex vivo. CD160 blocking antibody CL1-R2 was employed to clarify its role in NK cell mediated cytolysis. Then, the expression of CD160 on NK cells in peripheral blood from various patients with hematological malignancies were measured by flow cytometry.

RESULTSThe mRNA and protein levels of CD160 expressions on NK92 cells were confirmed by RT-PCR and Western blot, respectively. The flow cytometry results demonstrated that the strong positive expression of CD160 could be detected on the NK92 cell surface. NK92 could effectively kill K562 and THP-1 cells, while the cytolysis effect was abrogated in the presence of CD160 blocking antibody CL1-R2. The high levels of HVEM were expressed on both target cells, but the HLA class I molecules were absent on K562. The expression of CD160 on CD3CD56 NK cells in peripheral blood from patients with acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL) and allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients was significant lower than that in the normal controls (P<0.05).

CONCLUSIONThe cytolysis function of human NK cells is mediated partially by CD160 molecule. The decrease of CD160 expression on NK cells from patients with various hematological malignancies implies that down-regulation of CD160 expression may be a novel mechanism of tumor immune escape.

OBJECTIVE: To investigate the changes in autophagy of mesenchymal stem cells (MSCs) from patients with ankylosing spondylitis and explore the mechanism for decreased autophagy in ASMSCs. METHODS: MSCs collected from 14 patients with AS (ASMSCs) and from 15 healthy donors (HDMSCs) were cultured in the absence or presence of 25 ng/mL TNF-α for 6 h. Autophagy of the cells was determined by immunofluorescence staining of GFP-LC3B, and the results were confirmed by detecting the protein expressions of autophagy markers LC3 II/LC3 I and P62. The mRNA expressions of the related genes were detected using qRT-PCR, and the protein expressions of the autophagy markers and signaling pathway-related molecules were determined with Western blotting. TG100713 was used to block the PI3K/AKT/mTOR signal pathway, and its effect on autophagy of ASMSCs was evaluated. RESULTS: ASMSCs showed significantly weaker GFP-LC3B puncta staining and lower protein expression levels of LC3 II/LC3 I but higher levels of P62 protein (P < 0.05), indicating a decreased autophagy capacity as compared with HDMSCs. TNF-α-induced ASMSCs showed significantly higher protein expressions of p-PI3K/ PI3K, p-AKT/AKT and p-mTOR/mTOR than HDMSCs (P < 0.05), suggesting hyperactivation of the PI3K/AKT/mTOR signaling pathway in ASMSCs. Blocking PI3K/AKT/mTOR signaling with TG100713 eliminated the difference in TNF-α-induced autophagy between HDMSCs and ASMSCs. CONCLUSION In patients with AS, hyperactivation of the PI3K/AKT/mTOR signaling pathway results in decreased autophagy of the MSCs and potentially contributes to chronic inflammation.
Autophagy ; Humans ; Mesenchymal Stem Cells/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Signal Transduction ; Spondylitis, Ankylosing ; TOR Serine-Threonine Kinases/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*