高危人乳头瘤病毒58型E6基因的克隆及表达

目的：获得含人乳头瘤病毒(HPV)58型E6基因的克隆及表达重组体并体外表达E6蛋白．方法：聚合酶链方法从1例宫颈腺癌患者癌组织DNA中获得HPV58 E6基因,并将其与克隆载体pGEM—T Easy连接,获得重组体HPV58-E6-pGEM—T,继之以双酶切将E6基因与同样双酶切的线性化的pRSET-A表达载体连接,得到E6表达重组体pRSET-58E6,转化E．coliBL21(DE3),用LPTG诱导表达．结果：从1例宫颈癌患者中成功获得了少见HPV58型的E6基因并构建了其重组表达载体．经IPTG诱导后可表达M24000的6His HPV58E6融合蛋白,表达量占菌体蛋白的10％．结论：成功获得了少见HPV58高危型的E6基因,并可在E．coli中高效表达．     

Circular RNA FOXO3 accelerates glycolysis and improves cisplatin sensitivity in lung cancer cells via the miR-543/Foxo3 axis

Non-small cell lung cancer (NSCLC) is the most common cause of cancer-associated mortality worldwide. Our previous study revealed that circular RNA (circRNA)-FOXO3 is highly expressed in lung cancer and inhibits cell proliferation. However, to the best of our knowledge, at present, no study has focused on the specific mechanism of circRNA-FOXO3 in drug resistance. Therefore, the present study aimed to provide novel perspectives on the role of circRNA-FOXO3 in cisplatin (DDP) resistance in NSCLC. A Cell Counting Kit-8 assay was used to determine the viability of cells overexpressed with circRNA-FOXO3 and under DDP treatment. Glycolysis was analyzed by measuring glucose consumption and lactate production. The interaction of circRNA-FOXO3, microRNA 543 (miR-543) and Foxo3 was confirmed using a dual-luciferase reporter assay. It was revealed that circRNA-FOXO3 improved cell sensitivity to DDP and repressed glycolysis in DDP-sensitive and DDP-resistant NSCLC cells. Bioinformatics analysis, luciferase reporter assays, quantitative PCR and RNA pull-down assays were employed to verify the binding of circRNA-FOXO3 to miR-543. Functionally, inhibition of miR-543 could sensitize NSCLC cells to DDP, and overexpression of miR-543 at least partially abolished the circRNA-FOXO3-induced decrease in chemoresistance. Furthermore, it was revealed that Foxo3 was a direct target of miR-543. Notably, the inhibitory action of miR-543 silencing on DDP resistance and glycolysis was reversed by overexpression of Foxo3 in DDP-sensitive and DDP-resistant NSCLC cells. In conclusion, the present study demonstrated that circRNA-FOXO3 promoted DDP sensitivity in NSCLC cells by regulating the miR-543/Foxo3 axis-mediated glycolysis balance. The present findings may provide novel perspectives for the treatment of patients with NSCLC resistant to DDP.     

Bortezomib,Melphalan, and Prednisone With or Without Daratumumab in Transplant-ineligible Asian Patients With Newly Diagnosed Multiple Myeloma: The Phase 3 OCTANS Study

IntroductionIn the global phase 3 ALCYONE trial, daratumumab plus bortezomib/melphalan/prednisone (D-VMP) improved outcomes versus VMP in transplant-ineligible newly diagnosed multiple myeloma (NDMM) patients. Here, we report the primary analysis of the phase 3 OCTANS trial of D-VMP versus VMP in transplant-ineligible Asian NDMM patients.Patients and MethodsIn total, 220 patients were randomized (2:1) to receive 9 cycles of VMP (bortezomib 1.3 mg/m² subcutaneously twice weekly in Cycle 1 and weekly in Cycles 2 to 9; melphalan 9 mg/m² orally; and prednisone 60 mg/m² orally on Days 1 to 4 of each cycle) ± daratumumab 16 mg/kg intravenously weekly in Cycle 1, every 3 weeks in Cycles 2 to 9, and every 4 weeks thereafter until disease progression.ResultsAfter a median follow-up of 12.3 months, very good partial response or better rates (primary endpoint) were 74.0% versus 43.2% with D-VMP versus VMP (odds ratio, 3.57; 95% confidence interval [CI], 1.99-6.43; P < .0001). Median progression-free survival (PFS) with D-VMP versus VMP was not reached versus 18.2 months (hazard ratio, .43; 95% CI, .24-.77; P = .0033); 12-month PFS rates were 84.2% versus 64.6%. The most frequent grade 3/4 treatment-emergent adverse events with D-VMP/VMP were thrombocytopenia (46.5%/45.1%), neutropenia (39.6%/50.7%), and leukopenia (31.3%/36.6%).ConclusionD-VMP demonstrated a favorable benefit/risk profile in transplant-ineligible Asian NDMM patients. This trial was registered at www.ClinicalTrials.gov as #NCT03217812.     

完善医疗服务项目规范 强化医疗收费价格管理

2001年国家发展改革委、卫牛部和中医药管理局共同颁发了《全国医疗服务价格项目规范(试行)》(以下简称《项目规范》)文件,根据此规范陕西省制定了《陕西省医疗服务项目价格》(以下简称《项目价格》)并于2002年7月1日起正式在全省执行.