OBJECTIVE To investigate the effect and mechanism of gracillin from Reineckia carnea on autophagy in non- small cell lung cancer A549 cells. METHODS Using A549 cells as subjects， the effects of different concentrations of gracillin （0.25， 0.5， 1， 2， 4 μmol/L） on the proliferation of cells were detected by CCK-8 after being treated for different time （12， 24， 48 h）. Compared with the control group without medication， the effect of gracillin （2 μmol/L） on the formation of autophagosomes in cells was observed by transmission electron microscope after 24 h of exposure. The aggregation of GFP-LC3 on autophagosome membrane was detected by GFP-LC3 plasmid transfection after being treated with gracillin （0.25， 0.5， 1， 2 μmol/L） for 24 h. Quantitative real-time PCR and Western blot assay were used to detect the mRNA and protein expressions of family with sequence similarity 102 member A（FAM102A）， the expressions of autophagy-related proteins [p62， Beclin-1， microtubule-associated protein 1 light chain 3B （LC3B）]， and the expressions of phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway-related proteins in A549 cells after being treated with gracillin （0.25， 0.5， 1 and 2 μmol/L） for 24 h. RESULTS Gracillin significantly inhibited the proliferation of A549 cells in a concentration- and time-dependent manner. The IC50 was 2.55 μmol/L at 24 h. After 24 h of gracillin treatment， autophagosomes with bilayer membrane structure were found in the cell cytoplasm， and GFP-LC3 green fluorescent spots on autophagosome membrane were obvious， representing an increasing trend as drug concentration. Compared with the control group， mRNA and protein expressions of FAM102A （0.5， 1， 2 μmol/L groups）， protein expression of Beclin-1 （1， 2 μmol/L groups） and LC3B-Ⅱ/LC3B-Ⅰ ratio （2 μmol/L group） were significantly increased in different concentrations of gracillin groups， while the protein expression of p62 （1， 2 μmol/L groups）， and the protein phosphorylations of Akt （1， 2 μmol/L groups） and PI3K （2 μmol/L group） were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Gracillin can promote excessive autophagy in A549 cells by up-regulating mRNA and protein expressions of FAM102A and inhibiting PI3K/Akt signaling pathway， thus inhibiting cell proliferation.

Objective:To analyze the perioperative clinical characteristics of patients with oral cancer underwent radical resection and flap reconstrution and the nursing managements.Methods:From January 2020 to December 2020, 658 patients with oral cancer underwent radical resection and flap reconstrution in the Department of Oral and Maxillofacial Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University were retrospectively analyzed. All patients were divided into the elderly group (≥60 years) and the younger group (< 60 years), including 279 cases in the elderly group and 379 cases in the younger group. The perioperative clinical data of the patients were collected, and the perioperative general situation, postoperative complications and influencing factors of complications were analyzed.Results:There were differences between the two groups in the aspects of sex ( χ2 = 12.38, P<0.001), preoperative BMI ( t = 2.43, P = 0.015), smoking history ( χ2 = 18.34, P<0.001), preoperative anesthesia grade ( χ2 = 25.61, P = 0.001), preoperative coexisting disease ( χ2 = 46.97, P<0.001), whether oral floor or tongue cancer ( χ2 = 16.68, P<0.001), whether free flap ( χ2 = 6.81, P = 0.003), operation time ( t = 2.19, P = 0.029), preoperative test index hemoglobin ( t = 4.96, P<0.001), albumin ( t = 5.44, P<0.001), D-dimer( Z = -13.52, P<0.001), calcium levels ( t = 4.07, P<0.001) and postoperative complications ( χ2 = 14.55, P<0.001). Multivariate analysis of postoperative complications showed that the age ( OR = 1.021, 95% CI = 1.005-1.037, P = 0.011), preoperative D-dimer ( OR = 1.219, 95% CI = 1.026-1.447, P = 0.024) and the preoperative coexisting disease ( OR = 1.642, 95% CI = 1.108-2.432, P = 0.013) were the risk factors for the postoperative complications. Multivariate analysis of discharge with tube showed that the age ( OR = 1.017, 95% CI = 1.003-1.031, P = 0.017), preoperative BMI ( OR = 0.917, 95% CI = 0.873-0.963, P = 0.001), whether oral floor or tongue cancer ( OR = 2.135, 95% CI = 1.475-3.091, P<0.001), and operation time ( OR = 1.220, 95% CI = 1.120-1.328, P<0.001) were the related factors for the discharge with tube. Conclusion:In view of the above risk factors, it is beneficial for the physical and mental recovery of patients to pay attention to preoperative evaluation, make adequate preoperative preparation, postoperative nursing and observation, improve swallowing function training, prepare for discharge, pay attention to continuous nursing, and establish a tertiary hospital-community-family rehabilitation system.

We describe a case of spontaneous conception following ovarian stimulation, in which a singleton pregnancy was revealed by ultrasound at 17 gestational weeks, with a multi-cystic "honeycomb" pattern in part of the placenta. With close monitoring, the patient delivered a healthy male neonate through cesarean section at 38 gestational weeks. The clinical findings, combined with ultrasound, laboratory, pathological, and immunohistochemistry examination, and short tandem repeat genotyping, confirmed a twin pregnancy consisting of a complete mole and coexisting fetus. No obvious abnormalities were found in the mother or the boy during a four-and-a-half-year's follow-up.

Objective:To investigate the clinicopathological features, diagnosis and differential diagnosis of primary central nervous system T-cell lymphomas (TPCNSL), and to analyze its biological behavior and prognosis.Methods:Three cases of TPCNSL were collected from September 2014 to September 2019 in the First Affiliated Hospital of Nanjing Medical University. They were evaluated by HE, immunohistochemistry (IHC) and molecular genetics, and the relevant literature was reviewed.Results:Among the 6 816 brain tumors, 97 were primary central nervous system lymphomas (PCNSL), including 3 TPCNSL. There were two male and one female patients, aged 60, 67, and 82 years. Clinically, they were presented with varying degrees of limb numbness and unstable gait. Microscopically, the tumor cells were distributed diffusely or around blood vessels. They showed significant atypia and brisk mitotic activity. By IHC, they were positive for LCA, CD3, CD43, TIA-1, and perforin. Two of three cases were positive for CD5 and granzyme B. T-cell receptor gene rearrangement was clonal. EBER in situ hybridization was negative. The patients were followed for 1 to 6 months; one patient received chemotherapy and died of recurrence 3 months after surgery. One patient died of recurrence 5 months after operation alone. One patient remained recurrence and metastasis free more than 4 months post surgery.Conclusions:PCNSL is uncommon, and most are B-cell lymphomas, while T-cell lymphomas are even rarer. As the latter may show atypical clinical manifestations, diverse histologic morphology and poor prognosis, early diagnosis and timely treatment are particularly important for patients to improve survival.

Objective : To estimate the risk threshold of route index（RI）and mosq-ovitrap index（MOI）based on Breteau index（BI）,as supplements for dengue fever risk monitoring in specific habitats. Methods : Two towns and two streets were selected from nine towns（streets）in Jiashan County,and then one village（community）was selected from each of them as a Aedes albopictus monitoring site. The BI,RI and MOI were employed at the same time and area from April to October in the year 2018. Linear regression models were built with RI,MOI and BI to calculate the dengue risk threshold of RI and MOI according to BI. Results : The linear regression model of BI（X）and RI（Y）was Y=0.145+0.662X（P<0.05）,of BI（X）and MOI（Y）was Y=3.423+0.524X（P<0.05）. If BI=5（having risk of transmission of dengue fever）,then RI=3.455（95%CI：1.717-5.198）,MOI=6.043（95%CI：-0.327-12.417）. If BI=10（having risk of outbreak）,then RI=6.765（95%CI：5.018-8.518）,MOI=8.663（95%CI：2.260-15.071）. If BI=20（having risk of epidemic）,then RI=13.385（95%CI：11.326-15.453）,MOI=13.903（95%CI：6.352-21.461）. Conclusion The dengue fever risk threshold of RI estimated by BI had a narrow 95%CI and could be applied for dengue fever risk assessment,while the risk threshold of MOI had a wide 95%CI and the application value needed further study.

Objective To evaluate midterm outcomes of thoracic endovascular aortic repair (TEVAR) with in situ fenestration (ISF) to revascularize the aortic arch vessels.Methods From Feb 2012 to Dec 2014,10 patients underwent TEVAR with aortic arch vessels revascularized via ISF.There were 6 patients of thoracic aortic aneurysms (TAA) and 4 of type B aortic dissection (TBAD).Patients were followed for all-cause mortality,endoleak of post-TEVAR,integrity and patency of aortic endograft and branch vessels.Results Totally 11 branch vessels [10 left subclavian arteries (LSA),1 left common carotid artery (LCA)] via ISF were revascularized in 10 patients.Patients were followed-up for 24-55 mouths,mean of 42.80 months.1 TAA patient died in 2 years post-TEVAR unrelated to the operation.All fenestrations remained patent,and there were no endoleaks and no occlusion,compression,or fracture of stents.There were no postoperative strokes and left upper limbs ischemia.1 patient had distal aortic endograft pseudoaneurysms formation in 2 years post-TEVAR and underwent reTEVAR treatment.Conclusion Aortic arch vessels revascularization via ISF in TEVAR is safe and feasible.Midterm outcomes is satisfactory.

Objective To evaluate the outcome of the endovascular aortic repair (EVAR) of abdominal aortic aneurysms (AAA) with diabetes mellitus (DM) and analyze the factors that influence its mid-long-term curative effect.Methods From Jan 2004 to Jun 2014,follow-up data of 103 AAA patients with DM treated with EVAR was analyzed retrospectively.Results Effective follow-up visits were conducted on 88 patients (85.43％).The patients were tracked for an median of 3.59 years,with the longest follow-up 10.32 years.25 deaths occurred and the all-cause mortality rate was 24.3％.6 cases had intervention-related complications (5.8％),including endoleaks,thrombosis,embolism,infection,false aneurysms.2 secondary interventions were conducted on 2 patients.The five-year cumulative survival rate was 70％ and the ten-year cumulative survival rate was 61％.Conclusion EVAR is safe for AAA patients with DM.

Objective To study the safety and efficacy of detachable fibered coils occlusion system for visceral artery aneurysms.Methods Thirty-three visceral artery aneurysms patients (12 males and 21 females) with an average age of 63.8±3.5 years who underwent endovascular embolization with detachable fibered coils occlusion system combined with standard pushable coils in our hospital were included in this study.Of these patients,20 (60.6％),7 (21.2％),3 (9.1％),2 (6.1 ％),and 1 (3.0％) were respectively diagnosed with splenic artery aneurysm,renal aneurysm,superior mesenteric aneurysm,celiac aneurysm,and pancreaticoduodenal aneurysm.The mean diameter of aneurysms was 16-38 (24.0±3.4)mm.The success rate of detachable fibered coils occlusion system for visceral artery aneurysms,postoperative complications and surgical procedure time were recorded.The patients received CT or ultrasonographic follow up at months 3,6,12 and every year after operation.Results The success rate of detachable fibered coils occlusion system for visceral artery aneurysms was 100％.No intraoperative bleeding and coil migration occurred.One patient with splenic artery dissection did not undergo operation.The patients were followed up for 2-24 (12.5±2.3) months,during which no enlargement or rupture of visceral artery aneurysms and no operation-related severe complication or death occurred.Conclusion Detachable fibered coils occlusion system combined with standard pushable coils is safe and effective for mild and moderate visceral artery aneurysms.

Objective To explore the status quo of nursing practice of hemophilia-specialized nurses, to find out the shortcomings of current work and to provide the basis for promoting the development of hemophilia-specialized nursing in China. Methods The nursing staff attending the National Nursing Conference on Hemophilia in Shenzhen on November 7, 2015 were investigated with questionnaires. This investigation involved more than 40 hemophilia treatment centers nationwide. The nursing staff were required to fill out the Questionnaire for Hemophilia-Specialized Nurses developed by the Nursing Council, World Hemophilia Federation on the site, which were taken back once completed. Totally 54 questionnaires were handed out, and 43 valid questionnaires were taken back. Results Totally 55.8% of the nurses engaged in hemophilia nursing for less than 5 years, and 58.1% of them spent less than 10 hours on hemophilia nursing per week. And the number of hemophilia centers whose requirement for specialized nurses was a college degree accounted for 55.8%. The major nursing practice of China's hemophilia-specialized nurses included preliminary diagnosis-related content and self-injection training (>50%), and involved less specialized nursing practice, including consultancy, education, liaison, triage, physical examination, medication management, extended care, nursing research and clinical trial (20%-50%). The functions of management and medical advice accounted for<20%, and the least nursing practice was "school visit", accounting for only 4.7%. Merely 5 hemophilia centers carried out nursing practice items that accounted for over 50%. Nurses with different working years and education backgrounds were moderately correlated with the number of carried out nursing practice items. Conclusions The hemophilia-specialized nurses in China has an uneven level, and they need to expand the nursing work, do better in information registration and health education for hemophilia patients, improve the training proportion of family treatment, and meanwhile strengthen the publicity of hemophilia in elementary and middle schools.

Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.