Background Studies confirmed that hydroxycamptothecin cause the apoptosis of human Tenon capsule fibroblasts (HTFs) by protein kinase R-like endoplasmic reticulum stress kinase (PERK) single pathway.Autophagy and apoptosis are programmed cell death following stress reaction,so they remain a close association.However,the effect of hydroxycamptothecin on the autophagy of HTFs and its mechanism are still unclear.Objective This study was to explore the promoting effect of PERK signal pathway on hydroxycamptothecin inducing the autophagy of HTFs.Methods This study procedure was approval by Ethic Committee of Nanjing Medical University.Human Tenon capsule tissue was obtained from fresh adult donors.HTFs were cultured and passaged by explant-culture method and identified by immunofluorescence for vimentin and keratin.pLVX-PERK lentiviral packed by 293T cells was transfected into HTFs to obtain stable PERK-knockout cell line by puromycin selection.Then the HTFs were treated with 0.10 g/L of hydroxycamptothecin for 5 minutes and consecutively cultivated for 24 hours,and the untreated cells were used as the control group.Western blot assay was used to detect the expressions of autophagy specific proteins in the cells,including autophagy related gene 5 (ATG-5),Beclin-1,light chain 3 (LC-3).Cyto-ID staining was used to identify the autophagosome in the cells.The experimental results were analyzed and compared between different treating groups.Results The gray scales for the expressions of Beclin 1,ATG-5,LC-3-Ⅰ and LC-3-Ⅱ proteins in HTFs were 0.365:±0.045,0.765 ±0.055,0.120±0.030 and 0.215 ±0.035 in the control group,and those in the hydroxycamptothecin treated group were 0.980±0.070,1.495±0.095,0.585±0.025 and 0.785±0.055,showing a significant decline in the hydroxycamptothecin treated group(P=0.018,0.022,0.007,0.013).The green fluorescence of the autophagosome was stronger in the hydroxycamptothecin treated group compared with the control group.Western blot revealed that the gray scale of PERK expression in the cells was 0.130±0.030 in the PERK-knockout group,with a significant reduce in comparison with 0.765 ±0.055 of the control group (P =0.010).However,no obvious distinctions were seen in the band intensities of the expressions of Beclin-1,ATG-5 and LC-3 proteins between the two groups.Western blot indicated that the grey scale of the PERK expression in the cells was 1.790± 0.060 in the 0.10 g/L hydroxycamptothecin group,which was significantly higher than 0.880 ± 0.070 of the control group (P =0.010).Expression levels (gray scales) of Beclin-1,ATG-5,LC-3-Ⅰand LC-3-Ⅱ in the PERK-knockout+ 0.10 g/L hydroxycamptothecin group were 0.475 ± 0.045,0.390 ± 0.040,0.055 ± 0.015 and 0.075 ± 0.025,which were significantly lowed in comparison with 0.955 ± 0.065,0.765 ± 0.055,0.155 ± 0.015 and 0.280 ± 0.030 of the control+ 0.10 g/L hydroxycamptothecin group (P =0.026,0.031,0.042,0.034).In addition,the fluorescence intensity of autophagosomes was weaker in the PERK-knockout+0.10 g/L hydroxycamptothecin group compared with the control+0.10 g/L hydroxycamptothecin group.Conclusions Hydroxycamptothecin induces the autophagy of HTFs by PERK signal pathway.

Background The fibrosis of filtering area caused by proliferation of human Tenon fibroblasts (HTFs) is one of failure causes following glaucoma surgery.Researches revealed that hydroxycamptothecin can induce the apoptosis of HTFs,but its influence on autophagy of HTFs is unclear.Objective This study attempted to investigate whether hydroxycamptothecin can cause an alteration of autophagic activity in HTFs.Methods Human Tenon capsular tissue was obtained from 3 patients during strabismus correction surgery under the informed consent of patients and their parents for the primary culture and passaged of HTFs in DMEM containing 10％ fetal bovine serum.The generation 3 to 6 cells then were incubated with 0.0,0.5,1.0,4.0,10.0 mg/L hydroxycamptothecin for 24 hours,respectively.A cell counting kit-8 (CCK-8) was used to detect the cell viability in different treated groups.The autophagic activity of HTFs was evaluated by a Cyto-ID autophagy detection kit,and then the autophagic flux was evaluated by counting the Cyto-ID positive cells under a fluorescence microscope,and the green fluorescence intensity was determined by flow cytometry.Quantitative reverse transcriptase PCR (qRT-PCR) and Western blot analysis were employed to assay the relative expressions of autophagic-associated genes and their proteins in HTFs,including Beclin-1,autophagy related gene 5 (ATG-5) and light chain 3 (LC-3).Results The cell viability of HTFs in the 0.0,0.5,1.0,4.0 and 10.0 mg/L hydroxycamptothecin groups were (100.00 ± 6.44) ％,(91.70 ± 6.36) ％,(81.47 ± 6.00) ％,(68.43 ± 6.69) ％ and (59.97 ± 6.98) ％ respectively,showing a gradually declining trend with the increase of hydroxycamptothecin doses,with a significant difference among them (F=19.040,P＜0.001),and the viability of HTFs in the 1.0,4.0 and 10.0 mg/L hydroxycamptothecin groups were significantly decreased than the control group (P＜0.05,P＜0.01,P＜0.01).qRT-PCR analysis revealed that the relative expression levels of Beclin-1 mRNA,ATG-5 mRNA and LC-3 mRNA in 4.0 mg/L hydroxycamptothecin group were (3.225 ±0.346),(2.839 ±0.418) and (3.761±0.224) folds higher than those of the control group.The expressions of Beclin-1 and ATG-5 proteins were significantly increased in the 4.0 mg/L hydroxycamptothecin group in comparison with the control group,and the expression intensity ratio of LC-3-Ⅱ/Ⅰ was 0.965±0.159 in the hydroxycamptothecin group,which was significantly higher than 0.275 ±0.860 of the control group (P =0.003).Cyto-ID staining showed that the percentage of autophagic cells increased dramatically from (11.333±4.010) ％ to (55.000±9.013) ％ upon the exposure of HTFs to 4.0 mg/L hydroxycamptothecin (P=0.002).Flow cytometry analysis showed that the green fluorescence intensity in the 4.0 mg/L hydroxycamptothecin group was (3.037 ±0.513) fold relative to that in the control group,showing a significant difference between the two groups (P =0.003).Conclusions Hydroxycamptothecin can induce autophagy in HTFs in vitro.

Objective:To explore the role of hippocampal γ oscillation abnormality in sepsis-associated encephalopathy (SAE).Methods:Seventy male Sprague-Dawley rats (2-3 months) were randomly (random number) divided into three groups according to the random digital table method: sham, CLP, and CLP + dopamine 4 (D4) receptor agonists RO-10-5824 group. The SAE animal model was established by cecal ligation and puncture (CLP). On day 10-14 after surgery, the open field, novel object recognition, and fear conditioning tests were performed. After that, the hippocampus was collected to measure expressions of parvalbumin (PV) and D4 receptor. In another set of experiment, CA1 local field potential (LFP) were recorded, and the relationship between LFP and time with novel object was analyzed. Independent sample t-test was used for pairwise comparisons, and multiple comparisons were performed by one-way ANOVA, followed by the Tukey multiple comparisons test. Correlation was analyzed using Pearson correlation. Statistical significance was assumed when P<0.05. Results:Compared with the sham group, hippocampal PV (77.54±4.61)%, D4 expression (56.36±3.88)% and γ oscillation power (41.1±8.62)%, object exposure time (36±3) s, new object recognition rate (49±4)%, and scene stiffness time (56±7) s were decreased significantly ( P<0.05). However, RO-10-5824 treatment could increase hippocaml γ oscillation power (92.3±6.7)%, and reverse the decreased new object exposure time (44±3) s and new object recognition rate (63±4)%. Correlation analysis showed that hippocampal γ oscillation power was positively associated with new object exposure time ( r=0.609 2, P=0.015 9). There was no difference in total distance traveled or time spent in the center among groups ( P>0.05). Conclusion:Hippocampal γ oscillation abnormality might play a key role in cognitive impairment associated with SAE.

PURPOSE: Long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) has been implicated as an oncogene in the development and progression of osteosarcoma. This study aims to explore the mechanism of NEAT1 in osteosarcoma. MATERIALS AND METHODS: Expressions of NEAT1 and miR-194 in osteosarcoma tissues and cells were detected by quantitative real-time PCR. The effects of NEAT1 knockdown or miR-194 overexpression on cell proliferation, invasion, and apoptosis were determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide (MTT) assay, transwell invasive assay, and flow cytometry analysis, respectively. Luciferase reporter assay was performed to observe the possible interaction between NEAT1 and miR-194. RESULTS: NEAT1 was upregulated and miR-194 was downregulated in osteosarcoma tissues and cells. Knockdown of NEAT1 or overexpression of miR-194 suppressed proliferation and invasion and induced apoptosis of osteosarcoma cells in vitro. Luciferase reporter assay validated that NEAT1 could interact with miR-194 and negatively modulated its expression. Furthermore, inhibition of miR-194 reversed the suppression of proliferation and invasion and the promotion of apoptosis induced by NEAT1 depletion in osteosarcoma cells. CONCLUSION: Knockdown of NEAT1 suppressed proliferation and invasion and induced apoptosis in osteosarcoma cells by inhibiting miR-194 expression.
Apoptosis* ; Carcinogenesis ; Cell Proliferation ; Flow Cytometry ; In Vitro Techniques ; Luciferases ; Oncogenes ; Osteosarcoma* ; Real-Time Polymerase Chain Reaction ; RNA, Long Noncoding*

OBJECTIVE To study the in virto interaction of allicin combined with cefoperazone against clinical isolates of Pseudomonas aeruginosa.METHODS The protocol was designed by checkerboard method and the MICs of allicin combined with cefoperazone against the 17 strains of sensitive and 14 strains of drug-resistant P.aeruginosa were determined by broth dilution method,the FIC index was calculated according to MIC results.The combined effects were confirmed by FIC index.RESULTS The percentage of the FIC index less than 0.5,from 0.5 to 1,from 1 to 2,and more than 2 was 41.2-64.3% 35.7-41.2% 0-17.6%,and 0%,respectively.CONCLUSIONS Synergism and additivity of allicin combined with cefoperazone against P.aeruginosa are their main action,there are little autonomy and no antagonism.Allicin can significantly improve the antibacterial activity of cefoperazone against drug-resistant P.aeruginosa.

Objective To investigate the MRI features of solitary fibrous tumors(SFT)in the spinal canal.Methods MRI images of 5 cases with pathologically proved STF in the spinal canal were analyzed retrospectively.Results Of 5 lesions,there were 2 in the cervical spine,3 in the thoracic spine;1 in the epidural space and 4 in the subdural extramedullary space.On MRI plain scan,3 lesions showed homogeneous iso-intense signal on T1WI and hypo-intense signal on T2WI,2 lesions showed heterogeneous signal,1 showed patchy hypo-intense signal on T1WI and T2WI at the upper edge of lesion,which had been confirmed as hemorrhage and the other lesion showed internal cystic variation.All of the 5 lesions enhanced on enhancement scan,with moderate enhancement in 2 lesions and significant enhancement in 3 lesions.Cystic and hemorrhagic area were not enhanced.The"dural tail sign"was showed in 3 cases.Conclusion The diagnosis of SFT should be considered when a lesion shows a localized solitary mass in the spinal canal with hypo-intense on T2WI and moderate to significant enhancement.

Objective: Meta-analysis was used to compare the long-term efficacy and laryngeal function preservation rate of patients with advanced hypopharyngeal cancer treated with surgery plus radio(chemo)therapy (SRT) or non-surgery chemoradiotherapy (CRT). Methods: We searched publicly published articles on case-control studies of surgical and non-surgical comprehensive treatment of advanced hypopharyngeal cancer in PubMed, the Cochrane Library, Wanfang Database, Chinese Journal Full-text Database, and Chinese Science and Technology Periodical Database. The search language was limited to Chinese and English, and the period was from 1990 to 2018. These literatures were rigorously screened by inclusion and exclusion criteria. The data needed for this study were extracted and the Meta analysis was performed using RevMan 5.3 software. Results: A total of 13 literatures were included, and the overall quality of the literature was relatively high, and no significant publication bias was suggested. A total of 1 994 subjects, including 720 in the SRT group and 1 274 in the CRT group. The average 3-year overall survival rates were 42.9% in SRT group and 44.8% in CRT group,with no significant difference (OR=1.14, 95%CI: 0.62-2.06, P=0.68). The average 5-year overall survival rate (OR=1.42, 95%CI: 1.10-1.84, P<0.01), 5-year local recurrence-free survival rate (OR=1.68, 95%CI: 1.11-2.55, P=0.01) and 5-year local control rate (OR=2.17, 95%CI: 1.52-3.12, P<0.01) of SRT group were 46.4%, 47.4% and 71.2%, respectively, which were higher than those of non-surgical group (37.9%, 32.0%, and 52.2% respectively). The average laryngeal function preservation rate was 19.8%,being significantly lower than 80.6% of the non-surgical group(OR=0.03, 95%CI: 0.01-0.07, P<0.01). Conclusions SRT has better long-term efficacy, while CRT has better preservation of laryngeal function.

Lipids are important components of living organisms that participate in and regulate a variety of life activities. Lipids in plants also play important physiological functions in response to a variety of abiotic stresses (e.g. salt stress, drought stress, temperature stress). However, most research on lipids focused on animal cells and medical fields, while the functions of lipids in plants were overlooked. With the rapid development of "omics" technologies and biotechnology, the lipidomics has received much attention in recent years because it can reveal the composition and function of lipids in a deep and comprehensive way. This review summarizes the recent advances in the functions and classification of lipids, the development of lipidomics technology, and the responses of plant lipids against drought stress, salt stress and temperature stress. In addition, challenges and prospects were proposed for future lipidomics research and further exploration of the physiological functions of lipids in plant stress resistance.
Droughts ; Gene Expression Regulation, Plant ; Lipids ; Plants ; Stress, Physiological