Objective:To explore the mechanism by which miRNA-4469 (miR-4469) regulates the proliferation and invasion of renal cancer cells in vitro.Methods:The survival differences of patients with different expression levels of miR-4469 were analyzed based on the OncomiR database. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method was used to detect the expression of miR-4469 in renal cancer cell lines ACHN, OS-RC-2, SK-RC-20, 769-P, A498 and normal renal tubular epithelial cell line HK-2, and the renal cancer cells with the lowest expression level of miR-4469 were divided into miR-4469 group and control group, and were transfected with miR-4469 mimic and negative control sequence, respectively. The CCK-8 assay was used to detect the cell proliferation ability (expressed as absorbance value) in the two groups, and Transwell assay was used to analyze the number of invasive cells in the two groups. TargetScan Release 8.0 software was used to predict the binding site between miR-4469 and protein disulfide isomerase A4 (PDIA4) mRNA, and dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-4469 and PDIA4 mRNA. qRT-PCR method was used to detect the expression of PDIA4 mRNA in cells of each group, and Western blotting method was used to detect the expression levels of PDIA4 protein and PI3K-AKT-m-TOR pathway proteins in cells of each group.Results:Analysis of relevant data from the OncomiR database showed that compared with patients with low miR-4469 expression, the overall survival of renal cancer patients with high miR-4469 expression was better ( P < 0.001). The relative expression of miR-4469 in each renal cancer cell line was lower than that in HK-2 cells (all P < 0.05), and the expression of miR-4469 in 769-P cells was the lowest, which were selected to perform the subsequent experiments. The proliferation ability of 769-P cells in the miR-4469 group was lower than that in the control group ( P < 0.01). The number of 769-P cell invasions in the miR-4469 group were less than that in the control group [(19±3) cells vs. (64±7) cells, t = 5.44, P = 0.002]. Compared with the co-transfection of wild-type PDIA4 and miR-4469 negative sequence group, the relative luciferase activity of cells in the co-transfection of wild-type PDIA4 and miR-4469 mimic sequence group was lower (0.42±0.07 vs. 1.01±0.08, t = 5.74, P = 0.001); there was no statistical difference in cell luciferase activity between the co-transfected mutant PDIA4 and miR-4469 negative sequence group and the co-transfected mutant PDIA4 and miR-4469 mimic sequence group (0.99±0.11 vs. 1.02±0.11, t = 0.19, P = 0.001). The relative expression levels of PDIA4 mRNA in 769-P cells in the miR-4469 group were lower than that in the control group (0.98±0.23 vs. 7.19±2.23, t = 2.77, P = 0.032). Compared with the control group, the expression of PDIA4 protein and PI3K-AKT-m-TOR pathway-related p-PI3K, p-AKT, p-mTOR, and p-SGK1 proteins in 769-P cells in the miR-4469 group were all lower (all P < 0.05). Conclusions:miR-4469 may be related to the survival of renal cancer patients, and its expression is down-regulated in various renal cancer cell lines. miR-4469 may inhibit the proliferation and invasion of renal cancer 769-P cells by regulating the PI3K-AKT-m-TOR pathway through PDIA4.

Humans ; Adult ; Serum Albumin, Human ; Consensus ; Cardiac Surgical Procedures ; Thoracic Surgery

Objective:To explore the relationship between the relative expression of miRNA-676-3p and the survival of renal cancer patients, and its effect on the proliferation and invasion of renal cancer by targeting and regulating prefoldin 1 (PFDN1).Methods:OncoRank online software was selected to analyze the relationship between the relative expression of miRNA-676-3p and the survival rate of renal cancer patients. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the relative expression of miRNA-676-3p in renal cancer cell lines. Renal carcinoma CAKI1 cells were resuscitated, and the transfected miRNA-NC was used as the control group, and the transfected precursor miRNA-676-3p was used as the overexpression group. The relative expression of miRNA-676-3p was detected by RT-qPCR. The cell absorbance and invasion number of the two groups were measured by CCK-8 and Transwell invasion assays, respectively. The target gene of miRNA-676-3p was predicted and verified by referring to the TargetScan Release 8.0 website and dual-luciferase reporter gene experiment. The expression of PFDN1 gene and Wnt/β-catenin molecular pathway protein in the two groups of cells were determined by RT-qPCR and Western blotting, respectively. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The survival rate of renal cancer patients with high expression of miRNA-676-3p was significantly higher than that of renal cancer patients with low expression of miRNA-676-3p, the difference was statistically significant ( P<0.01). The relative expression of miRNA-676-3p in renal cancer cell lines was significantly lower than that in normal renal tubular epithelial cells, the difference was statistically significant ( P<0.01), and the relative expression of miRNA-676-3p in CAKI1 cells was the lowest, the difference was statistically significant ( P<0.01). The relative expression levels of miRNA-676-3p in the control and overexpression groups were 1.04±0.59 and 15.90±1.70, respectively, and the overexpression group was significantly higher than the control group, the difference was statistically significant ( P<0.01). After 24, 48, 60, and 72 h of culture, the absorbance of cells in the overexpression group was lower than that in the control group, the difference was statistically significant ( P<0.05). The number of invasion cells in the control group and the overexpression group were (115.90 ± 24.73) and (43.83 ± 21.94) cells, respectively, and the number of cell invasion in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). PFDN1 was the downstream target gene of miRNA-676-3p ( P<0.01). The relative expression of PFDN1 gene in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). The expression of Wnt/β-catenin molecular pathway proteins in the overexpression group was lower than that in the control group. Conclusions:Renal cancer patients with high expression of miRNA-676-3p had a higher survival rate. miRNA-676-3p inhibited the proliferation and invasion of renal cancer CAKI1 cells by significantly down-regulating the expression of PFDN1, thereby inhibiting the development of renal cancer.

Objective:The relative expression of lncRNA NPIPA9 in prostate cancer tissues was analyzed, and the relative expression of miR-210-3p and its effect on the growth and migration of prostate cancer cells were detected by overexpressing lncRNA NPIPA9.Methods:The relative expression of lncRNA NPIPA9 in prostate cancer tissues was analyzed by Oncomine database. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the relative expression of lncRNA NPIPA9 in prostate cancer cell lines DU-145, PC-3, C4-2B, 22Rv1, LNCaP and normal prostate epithelial cell RWPE-1. Prostate cancer PC-3 cells were cultured in vitro and divided into control group (transfected with control vector 100 nmol/L) and NPIPA9 group (transfected with lncRNA NPIPA9 vector 100 nmol/L). The proliferation activity of PC-3 cells was detected by CCK-8 method. The migration ability of PC-3 cells was detected by Transwell method. Potential target of lncRNA NPIPA9 were predicted using bioinformatics techniques. The dual-luciferase reporter gene assay determined the target binding relationship between lncRNA NPIPA9 and miR-210-3p. The effect of lncRNA NPIPA9 on the relative expression of miR-210-3p in prostate cancer cells was detected by RT-qPCR. The effect of lncRNA NPIPA9 on the expression of nuclear factor kappa-B (NF-κB) pathway proteins in prostate cancer cells was detected by Western blotting. Measurement data were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between two groups, one-way analysis of variance was used for comparison between multiple groups. Results:The expression of lncRNA NPIPA9 in prostate cancer tissue was lower than that in adjacent tissue, the difference was statistically significant ( P<0.01). The relative expression of lncRNA NPIPA9 in prostate cancer cell lines was lower than that in RWPE-1 cells, the difference was statistically significant ( P<0.01), and the relative expression of lncRNA NPIPA9 in prostate cancer PC-3 cells was the lowest, the difference was statistically significant ( P<0.01). Compared with the control group, lncRNA NPIPA9 had an inhibitory effect on the viability of prostate cancer PC-3 cells, the difference was statistically significant ( P<0.05). The migration numbers of PC-3 cells in the control group and NPIPA9 group were 101.70±8.63 and 45.97±8.83, respectively, and lncRNA NPIPA9 had an inhibitory effect on PC-3 cell migration, the difference was statistically significant ( P<0.01). lncRNA NPIPA9 can directly target miR-210-3p, the difference was statistically significant ( P<0.01). The relative expression of miR-210-3p in PC-3 cells in control group and NPIPA9 group were 5.32 ± 0.79 and 1.11 ± 0.56, respectively, and lncRNA NPIPA9 could directly down-regulate the expression of miR-210-3p in PC-3 cells, the difference was statistically significant ( P<0.01). Compared with the control group, lncRNA NPIPA9 can reduce the expression of NF-κB pathway proteins c-Myc, MMP-9, VEGF, p65, p50 in PC-3 cells. Conclusion:The expression of lncRNA NPIPA9 is down-regulated in prostate cancer tissues, and it reduces the proliferation and migration ability of prostate cancer PC-3 cells by targeting and negatively regulating miR-210-3p.

Objective:To explore the mechanism of Ma-Xing Shi-Gan decoction combined with Xiao-Chai-Hu decoction (hereinafter referred to as " Sanyang combined treatment" ) in alleviating colon injury in mice infected with influenza virus by transcriptome sequencing technique.Methods:The mouse model of colonic injury caused by influenza virus was induced by intranasal drip of influenza A virus H1N1 suspension. The mice were divided into Control group, Model group, and Sanyang combined treatment (SCT) group. Model group and SCT group were fed with PBS and Ma-Xing Shi-Gan decoction combined with Xiao-Chai-Hu decoction respectively. Seven days later, the colon tissues of each group were taken, the colon length and pathological damage were observed, and the transcriptome was sequenced to screen the significantly different genes between the SCT group and model group for Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis(GSEA).Results:After the therapy with SCT, the length of the colon of mice was significantly improved and the pathological injury of the colon was reduced. There are 92 differentially expressed genes between the SCT group and the model group. GO analysis indicated that the differential genes were enriched in biological processes such as regulation of cytokine and chemokine production, inflammatory response, defense response, immune response, regulation of NF-κB inducing kinase(NIK)/Nuclear factor-κB(NF-κB) signal and Mitogen-activated protein kinase(MAPK) cascade, as well as cell components related to intestinal barrier such as brush border membrane, brush border and microvilli. KEGG analysis indicated that the differential genes were enriched in Toll-like receptor signaling pathway, intestinal immune network for IgA production, complement and coagulation cascade, and Peroxisome proliferator-activated receptor(PPAR) signaling pathway. GSEA indicated that the intestinal immune network for IgA production, PPAR signaling pathway, propionic acid metabolism and butyrate metabolism were significantly up-regulated after the intervention with SCT, while apoptosis and MAPK signaling pathway were significantly down-regulated.Conclusions:Sanyang combined therapy can protect the intestinal tract of mice infected with influenza virus mainly through immunity, inflammation and metabolism pathways.

Objective:To explore the clinical and genetic characteristics of two children with a clinical diagnosis of Treacher Collins syndrome (TCS).Methods:Whole-exome sequencing was used to screen potential variants in the two children. Confirmation of suspected variants was performed through Sanger sequencing , multiplex ligation dependent probe amplification and real-time PCR in probands and their parents.Results:A heterozygous deletion variant, c. 4357_4360delGAAA, was detected in case one, while was de novo and verified by Sanger sequencing. Thevariant was classified as pathogenic(PVS1 + PM2+ PM6)according to ACMG guideline. The heterozygous deletion of exon 1-7 was seen in the same gene in case 2, which MLPA verified as heterozygous deletion of exon 1-6. This deletion was inherited from the father with a normal phenotype, and the father’s TCOF1 gene was suspected to be chimeric heterozygous deletion of exon 1-6 verified by MLPA. Conclusion:The identified variants in the TCOF1 gene probably underlie the two cases of TCS. There was no apparent correlation between genotype and phenotype. In addition, it shows a high interfamilial variability ranging from normal to full presentation of TCS. Genetic detection provided clinical diagnosis and genetic counselling for TCS patients .

Objective:To investigate the therapeutic mechanism of high-frequency repetitive transcranial magnetic stimulation (rTMS) in treating mild amnestic cognitive impairment (aMCI).Methods:Twenty-five patients with aMCI were randomly divided into an observation group of 13 and a control group of 12. The observation group was given 10-Hz rTMS over the left dorsolateral prefrontal cortex at 80% of the motor threshold-400 pulses a day, 5 times a week for 4 consecutive weeks. The control group received sham stimulation on the same schedule. Before and after the experiment, both groups were evaluated using the Montreal cognitive assessment (MoCA) and received fMRI scans.Results:After the intervention, the average MoCA score of the observation group had improved significantly more compared with that of the control group and compared with before the intervention. According to the fMRI results, regional homogeneity in the right middle frontal gyrus of the observation group had increased significantly, while that of the control group both there and in the left precuneus had decreased significantly.Conclusions:High-frequency rTMS can effectively improve the cognitive function of patients with aMCI and synchronize neuron activity in cognition-related brain regions.

Objective To explore the therapeutic effect of mild hypothermia on the inflammatory response, organ function and outcome in perioperative patients with acute Stanford type A aortic dissection (AAAD). Methods From February 2017 to February 2018, 56 patients with AAAD admitted in our department were enrolled and randomly allocated into two groups including a control group and an experimental group. After deep hypothermia circulatory arrest during operation, in the control group (n=28), the patients were rewarmed to normal body temperatures (36 to 37 centigrade degree), and which would be maintained for 24 hours after operation. While in the experimental group (n=28), the patients were rewarmed to mild hypothermia (34 to 35 centigrade degree), and the rest steps were the same to the control group. The thoracic drainage volume and the incidence of shivering at the first 24 hours after operation, inflammatory indicators and organ function during perioperation, and outcomes were compared between the two groups. There were 20 males and 8 females at age of 51.5±8.7 years in the control group, 24 males and 4 females at age of 53.3±11.2 years in the experimental group. Results There was no obvious difference in the basic information and operation information in patients between the two groups. Compared to the control group, at the 24th hour after operation, the level of peripheral blood matrix metalloproteinases (MMPs) was lower than that in the experimental group (P=0.008). In the experimental group, after operation, the awakening time was much shorter (P=0.008), the incidence of bloodstream infection was much lower (P=0.019). While the incidence of delirium, acute kidney injury (AKI), hepatic insufficiency, mechanical ventilation duration, intensive care unit (ICU) stays, or hospital mortality rate showed no statistical difference. And at the first 24 hours after operation, there was no difference in the thoracic drainage volume between the two groups, and no patient suffered from shivering. Conclusion The mild hypothermia therapy is able to shorten the awakening time and reduce the incidence of bloodstream infection after operation in the patients with AAAD, and does not cause the increase of thoracic drainage volume or shivering.

Objective To compare early outcomes of the minimally invasive aortic valve surgery (MIAVS) through right parasternal mini-thoracotomy with conventional mitral valve surgery (AVS),and evaluate feasibility and safety of MIAVS.Methods From January 2017 to December 2017,60 patients undergoing elective AVS in Nanjing First Hospital were prospectively enrolled in this study.There were 32 male and 28 female patients with their age of 28-72 (46.5 ± 10.2)years.Using a random number table,all the patients were randomly divided into a port-access MIAVS group (MIAVS group,n =20) and a conventional AVS group (conventional group,n =60).MIAVS group patients received port-access cardiopulmonary bypass (CPB) establishment via femoral artery,femoral vein and right internal jugular vein cannulation through right the 3rd in tercostal space with 5-6 cm right parasternal incision in length.Special MIAVS operative instruments were used for mitral valve repair or replacement.Conventional group patients received mitral valve repair or replacement under conventional CPB through median sternotomy.Perioperative clinical data,morbidity and mortality were compared between the 2 groups.Results There was no death in-hospital or shortly after discharge in this study.CPB time [(106.0 ± 21.0) minutes vs (73.0 ± 15.0) minutes] and aortic cross-clamping time [(78.0 ± 10.0) minutes vs (47.0 ± 7.0) minutes] of MIAVS group were significantly longer than those of conventional group (P ≤ 0.05).Postoperative mechanical ventilation time [(7.0 ±4.2) hours vs (10.2 ±5.3)hours],length of intensive care unit (ICU) stay [(19.0 ± 4.0) hours vs (27.5 ± 8.0) hours] and postoperative hospital stay [(8.5 ± 2.5) days vs (13.0 ± 3.0) days] of MIAVS group were significantly shorter than those of conventional group (P ≤ 0.05).Chest drainage volume within postoperative 12 hours [(100.0 ±40.0)ml vs (410.0 ±80.0)ml] and the percentage of patients receiving blood transfusion (15.0％ vs 55.0％) of MIAVS group were significantly lower than those of conventional group (P ≤0.05).Patients were followed up for 1-12 months,and the follow-up rate was 96.7％.There was no statistical difference in postoperative morbidity or mortality between the 2 groups (P ＞ 0.05).Conclusions Minimally invasive aortic valve surgery through right right parasternal mini-thoracotomy is a safe and feasible procedure for surgical treatment of mitral valve diseases.MIAVS can achieve similar clinical outcomes as conventional AVS,with more quickly recovery and less blood transfusion,and is a good alternative to conventional AVS.

Objective To investigate the impact of perioperative mild hypothermia on the neurological function and prog-nosis of patients with acute type A aortic dissection. Methods This study enrolled 65 patients with acute aortic dissection un-derwent surgery during the period of February 2017 to February 2018 and randomly divided them into mild hypothermia group and control group. After the process of deep hypothermic circulatory arrest,patients in the mild hypothermia group were re-warmed to 34 ℃ - 35 ℃ and maintained until 24 h after the operation. While,the patients in the control group were rewarmed to 36 ℃ and were treated with routine rewarm therapy. Baseline characteristics were recorded before the operation and neuro-logical and prognosis related indexes were recorded after the operation for all the patients. At the same time,peripheral venous bloods of all the patients were collected preoperatively and at 1、6、 12 and 24 h after the operation. Serum S 100β and neuron-specific enolase(NSE)levels were measured by ELISA kit. Results Compared with the control group,patients in the mild hypothermia group had a significantly shorter recovery time[ 10. 6 h(IQR:7. 6, 19. 1)vs. 25. 8 h(IQR: 13. 3,54. 2),P =0. 007]. At the same time,serum levels of NSE at 1 h and 6 h after operation and serum levels of S 100β levels at 1、6、 12 and 24 h after operation in the mild hypothermia group were significantly lower than those in the control group(P < 0. 05). In addi-tion,the length of stay in the mild hypothermia group was significantly shorter than that in the control group[ 19 days(IQR: 17, 23)vs. 24 days(IQR: 17,28),P = 0. 036]. However,there was no statistically difference in the incidence of delirium and cerebrovascular accidents between the two groups. Conclusion Perioperative mild hypothermia therapy can significantly re-duce brain cell damage in the patients with acute type A aortic dissection and can shorten postoperative recovery time and hospi-talization time,and thus improve the prognosis of patients.