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 investigate the effects of Bruner's constructivist theory on the critical thinking and practice performance of nursing students interning in internal medicine.Methods:One hundred and fifty-three undergraduate nursing students practicing in the department of internal medicine of a Beijing grade A tertiary hospital were selected by convenience sampling and divided into control group ( n=76) and experimental group ( n=77) according to the order of rotation time. The experimental group received teaching based on Bruner's constructivist theory, while the control group was given traditional teaching. SPSS 24.0 was used to perform the t test. Results:Compared with the control group, the experimental group showed a significantly higher critical thinking score [(306.13±33.00) vs. (325.03±32.09)], a significantly higher exit theory assessment score [(94.74±3.24) vs. (96.94±1.79)], a significantly higher exit skills assessment score [(86.68±11.95) vs. (90.23±9.17)], and a significantly higher degree of satisfaction with teaching [(180.08±13.35) vs. (187.91±14.50); all P<0.05]. Conclusions:Bruner's constructivist theory can help nurse students improve their critical thinking, theoretical and practical performance, and satisfaction with teaching, which enhances the effects of internal medicine nursing teaching.

Purpose: We aimed to evaluate whether the addition of pemetrexed is effective in improving progression-free survival (PFS) in epidermal growth factor receptor (EGFR)–mutated patients with or without concomitant alterations. Materials and Methods: This multicenter clinical trial was conducted in China from June 15, 2018, to May 31, 2019. A total of 92 non–small cell lung cancer (NSCLC) patients harboring EGFR-sensitive mutations were included and divided into concomitant and non-concomitant groups. Patients in each group were randomly treated with EGFR–tyrosine kinase inhibitor (TKI) monotherapy or EGFR-TKI combined with pemetrexed in a ratio of 1:1. PFS was recorded as the primary endpoint. Results: The overall median PFS of this cohort was 10.1 months. There were no significant differences in PFS between patients with and without concomitant and between patients received TKI monotherapy and TKI combined with pemetrexed (p=0.210 and p=0.085, respectively). Stratification analysis indicated that patients received TKI monotherapy had a significantly longer PFS in non-concomitant group than that in concomitant group (p=0.002). In concomitant group, patients received TKI combined with pemetrexed had a significantly longer PFS than patients received TKI monotherapy (p=0.013). Molecular dynamic analysis showed rapidly emerging EGFR T790M in patients received TKI monotherapy. EGFR mutation abundance decreased in patients received TKI combined chemotherapy, which supports better efficacy for a TKI combined chemotherapy as compared to TKI monotherapy. A good correlation between therapeutic efficacy and a change in circulating tumor DNA (ctDNA) status was found in 66% of patients, supporting the guiding role of ctDNA minimal residual disease (MRD) in NSCLC treatment. Conclusion EGFR-TKI monotherapy is applicable to EGFR-sensitive patients without concomitant alterations, while a TKI combined chemotherapy is applicable to EGFR-sensitive patients with concomitant alterations. CtDNA MRD may be a potential biomarker for predicting therapeutic efficacy.

Abstract To further standardize the procurement management of reagents and consumables in disease control and prevention institutions in Zhejiang Province and facilitate incorruptible health building, the procurement management for reagents and consumables in Zhejiang disease control and prevention institutions was created by Zhejiang Provincial Center for Disease Control and Prevention in 2019. Based on three-layer architecture of SaaS, PaaS and IaaS, this platform, which is easy to perform, safe and practical, provides modular portal website services, and its main functions include procurement budget management, procurement plan management, order management, bidding management, contract management, execution-of-contract and acceptance, and payment management. This platform, which was initiated to be run on early 2020, was found to be improve the procurement efficiency, safe management costs, reduce the internal control risk, and facilitate the containment of COVID-19, which may provide the support to improving procurement management and laboratory capability in disease control and prevention institutions.

Objective:To investigate the clinical, histologic, immunohistochemical (IHC) and molecular genetic features of clear cell carcinoma (CCC) of salivary gland in the head and neck regions.Methods:Seven cases of CCC diagnosed in the Department of Pathology, the First Affiliated Hospital of Nanjing Medical University from 2018 to 2021 were included. The clinical and pathologic data, HE sections and IHC staining were reviewed, and EWSR1 gene translocation was detected by fluorescence in situ hybridization (FISH). The relevant literature was also reviewed.Results:There were five males and two females, with an age range of 32 to 71 years (mean 50 years). The tumors were located in the palate, base of tongue, subglottic, right submaxillary and nasopharynx. Histologically the tumors were composed of sheets, nests, and trabecular of large, monomorphic cells which possessed abundant clear and eosinophilic cytoplasm. The stroma was characterized by abundant hyalinized fibrous strands admixed with cellular fibrous (desmoplastic) tissue. The tumor growth was infiltrative. IHC staining revealed positivity for CKpan and squamous cell immunophenotypic markers (CK5/6, p63 and p40), but negativity for myoepithelial markers (SMA, calponin, GFAP and CD10). The EWSR1 gene translocation was detected by FISH. The prognosis was excellent, with the follow-up periods ranging from 8 months to 33 months. During this period, six patients survived without tumor, only one patient with cervical lymph node metastasis.Conclusions:CCC of salivary gland is rare and needs to be differentiated from various other types of tumors containing clear cells. Awareness of the histopathologic characteristics, and combined with IHC and molecular genetic examination can avoid misdiagnosis. The biological behavior of the tumor is indolent with a good overall prognosis.

Objective:To investigate the clinicopathological features and differential diagnosis of polymorphous low-grade neuroepithelial tumor of the young (PLNTY).Methods:Five cases of PLNTY diagnosed at the First Affiliated Hospital and Affiliated Brain Hospital of Nanjing Medical University, Nanjing, China from 2019 to 2021 were collected. All cases were evaluated using clinical and imaging data, histology, immunohistochemical staining and molecular genetics. The relevant literature was reviewed.Results:There were two male and three female patients, aged 10 to 39 years, with an average age of 25 years. Clinically, the tumors were in the temporal lobe (3 cases), the lateral ventricle (1 case) and the left head of caudate nucleus (1 case). The clinical manifestations included epilepsy in 3 cases, right visual disturbance in 1 case, and post-trauma incidental finding in 1 case. Microscopically, the lesions were characterized with infiltrative growth, cellular pleomorphism (oligodendroglioma-like cells were always present, with low-grade, pleomorphic nuclei) and variable calcifications. Immunohistochemically, the tumor cells were positive for GFAP and Olig2. They also showed intense and diffuse expression of CD34 while CD34 expressing ramified neural elements were present in regional cortex. Ki-67 proliferation index was less than 3%. Molecular genetics showed the BRAF V600E mutation in 2 cases, the PAK5-Q337R missense mutation in 1 case, the FGFR2-CTNNA3 fusion in 1 case, and the FGFR2-INA and FGFR2-PPRC1 concomitant fusion in 1 case. No postoperative chemoradiotherapy was given. Follow-up intervals ranged from 3 to 29 months while no recurrence or metastasis was identified.Conclusions:PLNTY is uncommon. A definite diagnosis of PLNTY relies on histopathological examination and molecular genetics. It is important to distinguish PLNTY from high grade gliomas and avoid overtreatment. The recently reported the PAK5-Q337R missense mutation and the FGFR2-PPRC1 gene fusion in PLNTY may help diagnose and understand the pathogenesis of PLNTY.

Objective : To learn the situation of vaccine management in Zhejiang Province by SWOT analysis,so as to provide basis for normalization and standardization of vaccine management. Methods: Through supervision and investigation,the data of vaccine management related human resources and information construction of the CDCs in Zhejiang Province were collected. The comprehensive evaluation of vaccine management in Zhejiang Province was carried out by SWOT analysis,and the SWOT matrix was established for suggestions. Results: The advantages of vaccine management in Zhejiang Province lay in the standardized management of purchase and supply,the construction of vaccine management information system and the guarantee of cold chain system;the disadvantages lay in the insufficient allocation of human resources,new problems of cold chain management,the lack of standardized vaccine management mode and the incomplete coverage of vaccine and vaccination information system;the opportunities lay in the legalization of vaccine management,“Healthy Zhejiang”initiative and the “one run at most”reform in healthcare services;the threats lay in the shortage of vaccine,the gap between the expectation of the public on vaccine management and the current situation,and the reduced capacity of vaccine deployment in the provincial CDC. We should constantly improve the construction of vaccine management system,strengthen the standardized administration,regularly carry out risk assessment on vaccine management,accelerate the full coverage of vaccine and vaccination management information system,and explore the standard operation procedure of vaccine management in Zhejiang Province. Conclusions Although some achievements have been made in vaccine management in Zhejiang Province,there is still a gap from the goals of high-efficiency,high-level,normalization and standardization. The standardization and full coverage of the information system of vaccine management should be strengthen in the next stage.

Objective:To analyze and compare VP1 and VP4 genetic characteristics of human enterovirus 71(EV71) isolated from children with severe and mild hand-foot and mouth disease(HFMD).Methods:EV71 strains isolated from severe HFMD patients (6 cases) and mild HFMD patients (6 cases) in Taizhou district, Zhejiang province were included during 2016.Total virus RNA was extracted by Viral RNA Mini Extraction Kit, and reverse transcription polymerase chain reaction(RT-PCR) was used to amplify the sequence of the VP1 and VP4 genes of EV71.And then the sequencing results were compared with those of A, B, C genotype reference EV71 strains from GenBank by nucleotide alignment and amino acid alignment analysis.Results:There were no statistically significant differences in sex and age between the two groups(χ 2=14.51, t=2.82, all P<0.05). The homogeneity between EV71 strains from severe patients and mild patients was 95.8%~99.6% and 99.1%~100.0% for VP1 nucleotide sequences and amino acid sequences, respectively, while 95.0%~99.9% and 99.0%~100.0% for VP4 nucleotide sequences and amino acid sequences, respectively.The twelve EV71 strains isolated from HFMD patients in Taizhou shared the highest identity with EV71-genotype C, especially genotype C4a.In addition, compared with the mild patients, three respective strains from severe HFMD patients showed mutations at the residue 170 in the VP1 protein(V→L), residue 293 in the VP1 protein(A→S) and residue 7 in the VP4 protein(T→A). Conclusion:All EV71 strains isolated from severe and mild HFMD patients in Taizhou district share high homology of nucleotide and amino sequence, and all of them belong to subgenogroup C4a.The mutations in the VP1 and VP4 of EV71 might be related to HFMD disease severity.

Objective:To investigate dyslipidemia and relevant factors of AIDS patients in general hospital.Methods:From April 2017 to January 2018, 306 AIDS patients seen in Peking Union Medical College Hospital (PUMCH) were included, their general information such as demographic characteristics, underlying diseases, regimens for treatment and duration of antiretroviral therapy were recorded and dietary conditions were evaluated. General Self-Efficacy Scale (GSES), Social Support Rating Scale (SSRS), Internal Physical Activity Questionnaire (IPAQ) were adopted for the investigation.Results:Of the 306 patients, 179 had dyslipidemia. Univariate analysis suggested that singlehood, higher age and body mass index (BMI), proteinase inhibitor (PI) and non-nucleotide reverse transcriptase inhibitor (NNRTI) contained antiretroviral therapy (ART) (especially the former) were relevant risk factors. After multivariable analysis, higher age and BMI, high fat diet, high baseline CD4 counting and longer course of disease were related with dyslipidemia, whereas female and higher activity level may be associated with little benefit.Conclusions:Dyslipidemia was more common in AIDS patients than in general population, which were mainly due to the disease itself and ART regimens. Those patients were middle-aged or older, with high baseline CD4 counting, treated with PI and whose course of disease was more than two years should reserve more attention.

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.