Mass spectrometry analysis of intact protein N-glycosylation signatures of cells and sera in pancreatic adenocarcinomas / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Pancreatic cancer is among the most malignant cancers, and thus early intervention is the key to better survival outcomes. However, no methods have been derived that can reliably identify early precursors of development into malignancy. Therefore, it is urgent to discover early molecular changes during pancreatic tumorigenesis. As aberrant glycosylation is closely associated with cancer progression, numerous efforts have been made to mine glycosylation changes as biomarkers for diagnosis; however, detailed glycoproteomic information, especially site-specific N-glycosylation changes in pancreatic cancer with and without drug treatment, needs to be further explored. Herein, we used comprehensive solid-phase chemoenzymatic glycoproteomics to analyze glycans, glycosites, and intact glycopeptides in pancreatic cancer cells and patient sera. The profiling of N-glycans in cancer cells revealed an increase in the secreted glycoproteins from the primary tumor of MIA PaCa-2 cells, whereas human sera, which contain many secreted glycoproteins, had significant changes of glycans at their specific glycosites. These results indicated the potential role for tumor-specific glycosylation as disease biomarkers. We also found that AMG-510, a small molecule inhibitor against Kirsten rat sarcoma viral oncogene homolog (KRAS) G12C mutation, profoundly reduced the glycosylation level in MIA PaCa-2 cells, suggesting that KRAS plays a role in the cellular glycosylation process, and thus glycosylation inhibition contributes to the anti-tumor effect of AMG-510.

Staged breast reconstruction with a subpectoral silicone implant and autologous fat grafting after tissue expansion / 中华整形外科杂志

Objective@#To explore the clinical application of subpectoral silicone implant associated with autologous fat grafting after tissue expansion, for breast reconstructions.@*Methods@#From Jan 2013 to Dec 2016, a total of 15 female patients were admitted to the plastic surgery department of the First Affiliated hospital of Fujian Medical University. They were after or were prepared to unilateral modified radical mastectomy. Patients were aged 18-50 years old, with the average of 33.5 years. There were 8 cases of T1N0MO (T1micN0MO, n=1), 3 cases of T2N0MO, 1 case of T2N1MO, 1 case of T1N2MO, 1 case of T2N2MO and 1 case of T3N3MO. A tissue expander was inserted beneath the posterior pectoralis major in first surgery. The cavity stripped range is greater than the expander around 1cm. The lowest position of the expander was determined according to the IMF of the other breast. The tissue expander was gradually filled intraoperatively and postoperatively periodically, until the expander exceeds 30%-50% of the volume of the other breast. Three months later, as the soft tissue over the breast area has stretched enough, the expander was removed in a second operation and a permanent implant was inserted. Meanwhile, the autologous fat particles, harvested from thigh or abdomen, was centrifuged at 800×g for 3 minutes. Then it was injected into the subcutaneous and posterior pectoralis major space. And the other breast may be adjusted if necessary for symmetry.@*Results@#Fifteen patients was included in this study (immediately tissue expander implanted, n=8; delayed tissue expander implanted, n=7). All the expanders are round in shape. The interval between the two operations was 6-12 months, with an average of 7.4 months. The total salt-water solution injected was 310-450 ml per patient, with an average of 365 ml. The volume of breast implants was 180-280 ml. The injection volume of autologous fat was 65-230 ml, 122.7 ml on average. All incisions healed well. The follow-up period was 1-3 years, with an average of 2 years. The reconstructed breasts were in a natural shape, with a slight drop and bilateral symmetry. According to Harris breast shape evaluation criteria, all reconstructed breasts were graded as excellent.@*Conclusions@#The permanent implant associated with autologous fat grafting after tissue expansion is alternative for breast reconstruction.

Construction and identification of shRNA lentiviral vectors targeting histone lysine transferase KAT6B gene / 实用肿瘤学杂志

Objective The aims of this study were to construct short hairpin RNA(shRNA)lentiviral vector in breast cancer T47D cells,to carry out RNA interference on lysine acetyltransferase 6B(KAT6B/MORF)gene,to down-regulate its expression and to explore its function.Methods Two pairs of single-stranded short hairpin RNA(shRNA5 and shRNA8)and the corresponding control sequences(Scramble5 and Scramble8)were synthesized based on the CDS of KAT6B gene.Polymerase chain reaction(PCR) was used to amplify double-stranded and ligated with the entry vector(pENTR/pSM2(CMV)GFP),which were subjected to a doub-le digestion(EcoRl and Xhol)linearization and homologous recombination with the entry vector(pENTR/pSM2(CMV)GFP)to obtain an entry clone containing the desired fragment.The target fragment was recombined onto the target vector(pLenti x1 puro DEST)via the LR cloning reaction of the Gateway system.The lentiviral packaging plasmids were co-transfected into HEK-293T cells with two pairs of target plasmids. The supernatant of HEK -293T cells was collected and transformed into T47D cells. The expression of KAT6B protein was detected in T47D cells by Western blot.Results The single colony obtained from the transformation was identi-fied by sequencing,which was consistent with the target sequence,indicating that the lentiviral vector had been successfully construc-ted.The expression of KAT6B protein was significantly lower in the shRNA KAT6B group than that in the control group,which indica-ted that the constructed gene silencing vector could play a role in the KAT6B gene in T47D cells.Conclusion The shRNA lentiviral gene silencing vectors of KAT6B were constructed and identified in T47D cells,which indicated that the foundation for further study

The application of medpor and split-thickness skin graft in formation of cranioauricular sulcus during auricular reconstruction with Nagata method / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the application of medpor and split-thickness skin graft in formation of cranioauricular sulcus during auricular reconstruction with Nagata method.</p><p><b>METHODS</b>The first stage operation was fulfilled according to the Nagata two-stage method which involves fabrication and grafting of the costal cartilage framework. The second-stage ear elevation operation was undertaken 6 months later to form the cranioauricular sulcus. Split-thickness skin was taken from temporal and accipital area. After releasing the auricular framework and transplanting C shaped medpor at the rear side of framework, the temporaparietal fascia flap was transferred to cover postauricular medpor and framework. Then the split-thickness skin graft was implant on the fascia surface.</p><p><b>RESULTS</b>From July 2010 to August 2012, 20 cases (22 ears) were treated. Partial necrosis of temporaparietal fascia flap and framework exposure happened in 1 case. Successful ear reconstruction was achieved in other cases with satisfactory cranioauricular sulcus during the follow-up period of 6-18 months (average, 13 months).</p><p><b>CONCLUSIONS</b>The application of medpor and split-thickness skin graft in the ear elevation of Nagata method for auricular reconstruction for microtia can achieve satisfactory results. It not only avoids the obvious scar in the donor site due to harvesting full-thickness and intermediate-thickness skin, but also reduces chest trauma due to harvesting costal cartilage.</p>

Surgical treatment of Ebstein anomaly / 中国医师进修杂志

Objective To summarize the experience in surgical treatment of Ebstein anomaly,and evaluate the therapeutic effect.Methods Thirty-five patients of Ebstein anomaly were treated by operation.New York heart association(NYHA)heart function classⅠ?was in 11 cases,classⅡ?was in 14 cases,class III-Ⅳ?was in 10 cases.The cue of echocardiogram about tricuspid valve regurgitation in 24 cases were severe,5 cases were moderate and 6 cases were mild.Two patients accepted tricuspid valve replacement.Thirty-three cases were accepted tricuspid valve repair and right ventricle folded,in which 2 cases accepted Danielson method,and 31 cases accepted Carpentier method,among them,9 cases accepted one and a half ventricular repair.Associated heart anomaly was corrected at the same time.Results One patient died.There were 2 cases with third degree atrioventricular block after tricuspid valve replacement,2 cases with low cardiac output syndrome and 2 cases with atrial fibrillation combined premature ventricular contraction.The heart was lower by chest X-ray and echocardiogram examination,tricuspid valve regurgitation in 22 cases were disappeared,9 cases were mild,3 cases were moderate.All patients were followed up from 1 month to 7years,tricuspid valve regurgitation in 21 cases were disappeared,12 cases were mild,heart function were class Ⅰ-Ⅱ.Because of heart function aggravation,1 patient was operated again 3 years after operation.Condusions Ebstein anomaly is a rare congenital heart disease,Carpentier method tricuspid valve repair can decrease regurgitation obviously and protect right ventricular function.One and a half ventricular repair should be adopted according to the function of tricuspid valve and right ventricle.