Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common type of non-small cell lung cancer, and any change of miRNAs expression will affect the degree of target regulation, thus affecting intracellular homeostasis. This study verified that miR-186-5p could inhibit the proliferation, migration and invasion of LUAD cells by regulating PRKAA2. METHODS: Previous investigations found that the expression of miR-186-5p was markedly suppressed in LUAD. Bioinformatics method is used to predict the target protein related to ferroptosis downstream and inquire about its expression level in LUAD and its influence on the survival of patients. Double luciferase verified the binding site of PRKAA2 and miR-186-5p. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of PRKAA2. The effects of miR-186-5p of LUAD cells as well as the mechanism by which miR-186-5p inhibits Fer-1's sensitivity to ferroptosis were confirmed by EdU, Transwell, and scratch assays. The effect of miR-186-5p on the amount of reactive oxygen species (ROS) in LUAD cells was discovered using ROS experiment. Malondialdehyde (MDA) and glutathione (GSH) experiments were used to detect the effects of miR-186-5p and PRKAA2 on ferroptosis index of LUAD cells. The concentration of lipid ROS (L-ROS) in LUAD cells were measured using the L-ROS tests to determine the effects of miR-186-5p and PRKAA2. RESULTS: The expression of PRKAA2 is up-regulated, and a high level of PRKAA2 expression was associated with a poor prognosis for patients with LUAD. Overexpression of miR-186-5p decreased the gene and protein expression of PRKAA2. By promoting ferroptosis, miR-186-5p overexpression prevented lung cancer cells from proliferating, invading, and migrating. ROS could be produced in higher amounts in LUAD cells due to miR-186-5p. Overexpression of miR-186-5p and knockdown PRKAA2 up-regulated MDA content and reduced GSH content in LUAD cells, respectively. miR-186-5p could increase the content of L-ROS and promote the ferroptosis sensitivity of LUAD cells by targeting PRKAA2. CONCLUSIONS miR-186-5p promotes ferroptosis of LUAD cells through targeted regulation of PRKAA2, thus inhibiting the proliferation, invasion and migration of LUAD.
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Humans ; Lung Neoplasms/genetics* ; Carcinoma, Non-Small-Cell Lung ; Ferroptosis/genetics* ; Reactive Oxygen Species ; Adenocarcinoma of Lung/genetics* ; MicroRNAs/genetics* ; 3,4-Methylenedioxyamphetamine ; Cell Proliferation/genetics* ; Cell Movement/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor ; AMP-Activated Protein Kinases

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common clinical histological subtype of lung cancer and microRNAs (miRNAs) are a type of small non-coding RNAs which play a central role in cells. miR-30b-3p plays a key effect in many types of carcinoma, but there is still very little research on how it works in lung adenocarcinoma. The role and mechanism of miR-30b-3p in the proliferation and invasion of LUAD were explored in this study, to provide new targets for inhibiting the proliferation and invasion of LUAD. METHODS: NCBI database was used to screen out miRNA with obvious differential expression, and the differential expression and survival curve were searched by StarBase database. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression of miR-30b-3p in each lung adenocarcinoma cell line. 5-ethynyl-2'-deoxyuridine (EdU) cell proliferation assay and Transwell invasion assay were used to detect the proliferation and invasion of A549 cells in each group. The target genes of miR-30b-3p were determined by the target gene prediction websites. Western blot assay was used to detect the expression of COX6B1 in each group of A549 cells. Double luciferase assay was used to verify the targeted binding relationship between miR-30b-3p and COX6B1. RESULTS: The expression of miR-30b-3p in lung adenocarcinoma tissues and lung adenocarcinoma cells was downregulated (P<0.05). Low expression levels of miR-30b-3p were associated with poor prognosis in patients with lung adenocarcinoma (P=0.005,8). Overexpression of miR-30b-3p could inhibit the proliferation and the invasion of lung adenocarcinoma cells (P<0.05). Double luciferase assay proved that miR-30b-3p could target and bind to COX6B1 (P<0.05). Western blot analysis showed that the overexpression of miR-30b-3p could downregulate the expression of COX6B1 in A549 cells (P<0.05). EdU cell proliferation assay and Transwell invasion assay showed that the overexpression of miR-30b-3p could reverse the promoting effect of upregulation of COX6B1 on proliferation and invasion in lung adenocarcinoma cells (P<0.05). CONCLUSIONS miR-30b-3p acts as a tumor suppressor gene in lung adenocarcinoma, and it can inhibit the proliferation and invasion of lung adenocarcinoma by targeting the expression of COX6B1.
Adenocarcinoma of Lung/pathology* ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms/pathology* ; MicroRNAs/metabolism*

OBJECTIVE: To prepare zedoary turmeric oil compound liposomes (ZTOC-LPS) and evaluate its quality.METHODS: The preparation method of liposome, the addition amount of soybean phosphatidylcholine (SPC), ratio of SPC to cholesterol (CH) in lipid, drug-lipid ratio of zedoary turmeric oil (ZTO), drug-lipid ratio of tretinoin in formulation, and water bath temperature were screened using encapsulation efficiency and drug-loading amount of ZTO (represented by germacrone) and tretinoin as investigation indexes. Quality evaluation and primary stability investigation were conducted for liposomes prepared by optimal preparation technology. RESULTS: The optimal preparation method was ethanol injection method; The optimal preparation technology were SPC 4 mg in 1 mL lipid, the mass ratio of SPC to CH 3:1, the ratio of ZTO to lipid 1:9, the ratio of tretinoin to lipid 1:70, water bath temperature of 55 ℃. Encapsulation efficiencies of ZTO and tretinoin were (64. 63 ± 1. 00)％ and (90. 33 土 0. 72)％ in 3 batches of ZTOC-LPS, respectively. Drug-loading amount of ZTO and tretinoin were (9. 09 ± 0. 14)％ and (1. 43 ± 0. 02)％, respectively. Particle size was (257. 41 ± 7. 58) nm, Zeta potential was (-38. 77 ± 0. 81) mV,PDI was 0. 10 ± 0. 04; the results of centrifugal acceleration test showed that the liposomes had good physical stability. No obvious change was found in each investigation index of ZTOC-LPS that stored at (4 ± 2) ℃ for 1 month. CONCLUSIONS: Established preparation technology is simple and feasible, the quality of the prepared ZTOC-LPS conforms to the requirements, and it can provide reference for the following research of ZTOC-LPS.

OBJECTIVETo investigate the effect of curcumin against cigarette smoke extract (CSE)- induced oxidative stress in human bronchial epithelial cells and explore the underlying mechanism.

METHODSHuman bronchial epithelial cell line 16HBE was treated for 24 h with curcumin, CSE, CSE + curcumin, and CSE + curcumin with transfection by a short hairpin RNA targeting PPARγ (shPPARγ). MTT assay was used to observe the changes in the cell viability after the treatments. Quantitative real-time PCR was performed to detect the mRNA expressions of tumor necrosis factor- (TNF-), iNOS and PPARγ in the cells, and the protein expressions of iNOS, PPARγ and the phosphorylation of NF-κB p65 were detected using Western blotting.

RESULTSThe treatments did not cause significant changes in the cell viability. Exposure to CSE for 24 h significantly lowered PPARγ expression and increased TNF- and iNOS expressions and phosphorylation of NF-κB p65 in the cells. The effects of CSE were significantly suppressed by curcumin, but transfection of the cells with shRNA-PPARγ obviously abrogated the suppressive effects of curcumin.

CONCLUSIONSCurcumin suppresses CSE-induced oxidative stress and inflammation via the PPARγ/NF-κB signaling pathway in 16HBE cells, suggesting the potential of curcumin in the treatment of chronic obstructive pulmonary disease.

Objective To investigate the clinical efficacy and safety of visual standard channel combined with visual ultrafine channel PCNL precision puncture in treatment of complex renal calculi. Methods From June 2015 to October 2016, 48 cases of complicated renal calculi were treated with multi-channel lithotripsy with visual standard channel ultrasonic pneumatic lithotripsy combined with visual superfine channel PCNL precision puncture holmium laser lithotripsy. Including 10 cases of staghorn stone, 38 cases of multiple renal stones. Results 110 channels were established in 48 patients. 4 cases of preoperative renal insufficiency with infection in the puncture found in the pus and stones load larger, intraoperative diarrhea and PCNL simple treatment of obstruction site stones; 44 cases to complete one of the surgery: There were single channel established in every one of 5 cases, and double channels established in every one of 24 cases, three channels in established in every one of 15 cases; There were two cases of surgery in 8 cases and there were 12 new channels established. The average time of unilateral first operation was 75 (35 ~ 125) min. The first clearance rate was 79.2% (38/48), and the total clearance rate of postoperative stone was 87.5% (42/48). 6 cases of residual stone combined with ESWL and drug row of stone, followed up for 3 months, 6 cases of stone row net, the total stone clearance rate of 100.0% (48/48). Two consecutive postoperative no sepsis, bleeding, ureteral injury and other serious complications. Conclusions Visual standard channel combined with visual superfine channel PCNL precise puncture for the treatment of complex renal calculi is safe and effective, with high fruiting rate and low complication, which can be popularized in clinical practice.

Objective The objectives of this study were to compare the clinicopathological features of different Lauren type gastric cancer,to carry out survival analysis,and to screen the prognostic factors.Methods The clinical pathologic data of gastric cancer patients who underwent surgical treatment at the Affiliated Tumor Hospital of Harbin Medical University from January 1,2007 to June 30,2007 were retrospectively analyzed.The 633 cases of gastric cancer patients were divided into intestinal type gastric cancer and diffuse type of gastric cancer groups,which were analyzed clinical and pathological characteristics and survival.Results Compared with diffuse type of gastric cancer,the proportion of intestinal type gastric cancer was slightly high(51.66% vs. 48.34%),the proportion of male was also high(2.94:1 vs.2.03:1,P=0.035).These could be more often in elderly patients age(≥60 years)(54.43% vs.35.94%,P<0.001).The prognosis of intestinal type gastric cancer was significantly better than diffuse gastric cancer(median survival time:90.9 months vs.37.33 months, P=0.014).Multivariate analysis showed that age ≥60 years old,CA199 abnormalities,larger tumors,poor differ-entiation,serosal invasion,initial lymph node metastasis,palliative surgery and non-pyloric resection were poor prognostic factors in gastric cancer.Conclusion Lauren type can be better respond to the clinicopathological fea-tures of different gastric cancer and guide to the prognosis.

Objective To explore the feasibility and safety of visualization puncture combined with flexible ureteroscopy in the treatment of lower calyx stones.Method Visualization puncture combined with flexible ureteroscopy to treat the lower calyx stones was done in our center from January to August 2016 in our hospital.32 cases of patients were enrolled to have a retrospective analysis.There were 18 males and 14 females,aged from 25 to 65 years,with an average age of 43 years.The diameter of stone was 1.0-2.0 cm,with an average of (1.4 ± 0.6) cm.We used general anesthesia and then adjusted the surgery bed to operation side lateral elevation was 30 °-35.Flexible ureteroscopy with 200μm holmium laser was used firstly to break calculi as much as possible.Ultrasound-guided F4.8 visualization puncture system was used to establish F4.8 channel.The power option was 2001μm hohnium laser to crush calculus of the renal calculi to treat the calculus of the distal end of soft lens which still can not be touched by ureteroscopy.Routine nephrostomy tube was not placed.The soft ureter sheath F5 double-J tube,and indwelling balloon catheter were routinely placed.We removed the catheter after 1-2 days and the double J tube after 4 to 6 weeks.Results The flexible ureteroscopy lithotripsy operation time was 8-25 mins in all of the 32 patients.Visualization puncture channels were successfully established in 3-7 mins,and the visualized puncture stone search rate of 100％ (32/32).The success rate of first stage lithotripsy was 93.8％ (30/32).Two cases of lower calyx stones diverticulum diverted to PNCL due to poor visibility by bleeding.The operation time was 30-60 mins and the average of 45 mins.KUB review at day one after the surgery showed that there were residual stones in 5 cases.The stone free rate at one month after the surgery is 100.0％.The average postoperative hospital stay was (2.0 ± 1.5) days.There were uo bleeding,ureteral avulsion and perforation,septic shock,pleural effusion and intestinal injury and other serious complications.Conclusions Navigation ultrasound-guided visualization puncture combined with flexible ureteroscopy is safe and effective to treat lower calyx stones.

Objective To analyze the relationship between Arg -1 expression and the clinical pathologi-cal factors ,proliferation and prognosis value in patients with colorectal cancer .Methods The expression of Arg-1 was observed in normal tissues ,chronic inflammatory tissues ,and adenomas inflammatory carcinoma tissues of mice.At the same time,Arg-1 expression was observed in human colorectal cancer adjacent tissues ,inflamed tis-sues and colorectal cancer tissues .Arg-1 expressed in 20 cases colorectal inflammation -cancer model in mice . Arg-1 expressed in 20 normal colorectal tissues .Fiftheen colitis tissues and 110 colorectal cancer tissues were examined by Immunohistochemistry .Statistical analysis was used to analyze the changes of Arg -1 expression in different groups of mice and human colon tissue cases .Results Arg-1 protein expression in normal tissues of mice was gradually increased in colon ,chronic inflammatory tissues,adenomas,inflammatory carcinoma,with sta-tistical significances(P<0.05).Arg-1 expression in para -carcinoma tissue,colitis tissues,colorectal cancer tissues was gradually increased with statistical significance (P<0.05).Conclusion Arg-1 protein is associat-ed with colorectal cancer TNM stage .Arg-1 protein may be involved in occurrence and development process of inflammation-associated colon tumor and may be a candidate of proliferated and prognostic biomarker in patients with colorectal cancer .