Thrombin promotes epithelial ovarian cancer cell invasion by inducing epithelial-mesenchymal transition

Yi-Cun Zhong,Ting Zhang,Wen Di,Wei-Ping Li 대한부인종양학회 2013 Journal of Gynecologic Oncology Vol.24 No.3

Objective: Over-expression of thrombin in ovarian cancer cells is associated with poor prognosis. In this study, we investigated the role of thrombin in inducing epithelial-mesenchymal transition (EMT) in SKOV3 epithelial ovarian cancer cells. Methods: After thrombin treatment SKOV3 cells were subjected to western blots, reverse-transcription PCR, and enzyme-linked immunosorbent assay to quantify EMT-related proteins, mRNA expression of SMAD2, DKK1, and sFRP1, and the secretion of matrix metalloproteinases (MMPs) and cytokines. Meanwhile, invasion ability was evaluated using transwell assays. Results: The results indicated a dose- and time-dependent down-regulation of E-cadherin and upregulation of N-cadherin and vimentin in thrombin-treated SKOV3 cells, compared with the thrombin-free control group (p<0.05). There was a dose-and time-dependent increase in the levels of SMAD2 and DKK1 mRNAs and a decrease in the levels of sFRP1 mRNA in thrombin-treated SKOV3 cells compared to control cells (p<0.05). Thrombin-treated SKOV3 cells exhibited increased secretion of MMP-9, MMP-2, interleukin (IL)-8, and IL-6 and increased invasion compared to untreated cells (p<0.05). Thrombin altered the morphology of SKOV3 cells to a spindle-like phenotype. Addition of hirudin to thrombin-treated cells reversed the effects of thrombin. Conclusion: Thrombin induced EMT and promoted the invasion of SKOV3 cells, possibly via distinct signaling pathways. Hirudin inhibited the effects of thrombin, suggesting that anticoagulant therapy could be a novel therapeutic strategy for ovarian carcinoma.

Expression of Fas/FasL in CD8⁺ T and CD3⁺ Foxp3⁺ Treg Cells - Relationship with Apoptosis of Circulating CD8⁺ T Cells in Hepatocellular Carcinoma Patients

Guo, Cun-Li,Yang, Xiu-Hua,Cheng, Wen,Xu, Yi,Li, Jie-Bing,Sun, Yi-Xin,Bi, Yu-Mei,Zhang, Lei,Wang, Qiu-Cheng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Aims: Dysfunction of the host immune system in cancer patients can be due to a number of factors, including lymphocyte apoptosis. Several studies showed that $Foxp3^+T$ cells take part in inducing this process by expressing FasL in tumor patients. However, the relationship between apoptosis, $CD8^+T$ cells and $Foxp3^+T$ cells in HCC patients is still unclear. The present study was designed to investigate the correlation between apoptosis levels and Fas/FasL expression in $CD8^+T$ lymphocytes and $Foxp3^+T$ cells in patients with HCC. Methods: $CD8^+T$ cells and $CD3^+Foxp3^+T$ cells were tested from peripheral blood of HCC patients and normal controls and subjected to multicolor flow cytometry. The expression of an apoptosis marker (annexin V) and the death receptor Fas in $CD8^+T$ cells and FasL in $CD3^+Foxp3^+T$ cells were evaluated. Serum TGF-${\beta}1$ levels in patients with HCC were measured by enzyme-linked immunosorbent assay. The relationship between apoptosis and Fas expression, as well as FasL expression in $CD3^+Foxp3^+T$ cells was then evaluated. Results: The frequency of $CD8^+T$ cells binding annexin V and Fas expression in $CD8^+T$ cells, were all higher in HCC patients than normal controls and the proportion of apoptotic $CD8^+T$ cells correlated with their Fas expression. Serum TGF-${\beta}1$ levels correlated inversely with $CD3^+Foxp3^+T$ cells. Conclusions: Fas/FasL interactions might lead to excessive turnover of $CD8^+T$ cells and reduce anti-tumor immune responses in patients with HCC. Further investigations of apoptosis induction in $Fas^+CD8^+T$ cells in vitro are required.

Molecular characterization, activity analysis and transcriptional detection of chitinases encoded in the genome of Spodoptera exigua

Lei Hea,Huan Yu,Cun-Yi Xu,Ying Zhao,Fu-Xiang Yang,Ya-Dong Guo,Guo-Hua Huang 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.2

Insect chitinases are necessary hydrolytic enzymes for chitin degradation, insect molting and metamorphosis. In this study, five chitinases encoded in the genome of Spodoptera exigua (SeCHIT7, SeCHIT11, SeCHIT12, SeCHIT13 and SeCHIT14) were identified by reverse transcription PCR. Phylogenetic analysis indicated that SeCHIT7 belonged to Group III, and SeCHIT11/12/13/14 belonged to Group VIII. Real-time quantitative PCR analyses showed that SeCHIT7 had an extensive transcription at the fourth- and fifth-instar larval and pupal stages, while SeCHIT11 had the highest transcription level at the egg stage, and the transcription of SeCHIT12 increased by over 1000 times from pre-pupal to pupal stage. Tissue-specific analyses showed that these three SeCHITs (SeCHIT7, SeCHIT11 and SeCHIT12) were mainly transcribed in the midgut and fat body. Chitinase activity assays suggested that the activity had a lower level at the egg stage and peaked at the pupal stage. The activities were increased by 9.4 times from egg to first-instar larval stage. Tissue specificity analysis showed that the highest activity was observed in the fat body, followed by hemolymph and cuticle of the pre-pupal stage. Overall, these results provided valid information for further research on the biological function and regulation of chitinases in S. exigua.

Effect of Microstructure on Mechanical Property of Carbon Steel/Strainless Steel Rolling Clad Plate

Li Hai-bin,Zhou Cun-long,Fan Xiao-ning,Kong Yi-gang,Huang Qing-xue,Ma Qin 보안공학연구지원센터 2016 International Journal of u- and e- Service, Scienc Vol.9 No.11

The bonding interface microstructure and mechanical property of clad plate with different hot rolling schedules were researched. The results show that the carbide layer thickness of bonding interface increases firstly and then decreases with increasing reduction ratio. The carbide layer of bonding interface is the major factor relating to tensile strength, so the experimental values of clad plate are slightly higher than the calculated ones. Moreover, the bending strength of clad plate is mainly influenced by the thickness ratio and the carbide quantity of SS, while the carbide layer thickness of bonding interface is an important factor affects the bending strength.

Trans-Membrane Transport of n-Octadecane by Pseudomonas sp. DG17

Fei Hua,Hong Qi Wang,Yi Li,Yi Cun Zhao 한국미생물학회 2013 The journal of microbiology Vol.51 No.6

The trans-membrane transport of hydrocarbons is an important and complex aspect of the process of biodegradation of hydrocarbons by microorganisms. The mechanism of transport of 14C n-octadecane by Pseudomonas sp. DG17,an alkane-degrading bacterium, was studied by the addition of ATP inhibitors and different substrate concentrations. When the concentration of n-octadecane was higher than 4.54 μmol/L, the transport of 14C n-octadecane was driven by a facilitated passive mechanism following the intra/extra substrate concentration gradient. However, when the cells were grown with a low concentration of the substrate, the cellular accumulation of n-octadecane, an energy-dependent process, was dramatically decreased by the presence of ATP inhibitors, and n-octadecane accumulation continually increased against its concentration gradient. Furthermore, the presence of non-labeled alkanes blocked 14C n-octadecane transport only in the induced cells, and the trans-membrane transport of n-octadecane was specific with an apparent dissociation constant Kt of 11.27 μmol/L and Vmax of 0.96μmol/min/mg protein. The results indicated that the transmembrane transport of n-octadecane by Pseudomonas sp. DG17 was related to the substrate concentration and ATP.

Predicting the surgical reparability of large-to-massive rotator cuff tears by B-mode ultrasonography: a cross-sectional study

Po-Cheng Chen,Kuan-Ting Wu,Yi-Cun Chen,Yu-Chi Huang,Ching-Di Chang,Wei-Che Lin,Wen-Yi Chou 대한초음파의학회 2022 ULTRASONOGRAPHY Vol.41 No.1

Purpose: This study aimed to compare the ability of B-mode ultrasonography and magnetic resonance imaging (MRI) to predict the repairability of large-to-massive rotator cuff tears (RCTs). Methods: This cross-sectional study included participants with large-to-massive RCTs who underwent arthroscopic repair. B-mode ultrasonography and MRI were conducted prior to arthroscopic repair. B-mode ultrasonography was used to evaluate the echogenicity of the rotator cuff muscle using the Heckmatt scale. Intra-rater and inter-rater reliabilities were examined for two independent physicians. MRI was used to evaluate the degrees of tendon retraction, fatty infiltration of rotator cuff muscles, and muscle atrophy. Finally, two experienced orthopedic surgeons performed surgery and decided whether the torn stump could be completely repaired intraoperatively. Results: Fifty participants were included, and 32 complete repairs and 18 partial repairs were performed. B-mode ultrasonography showed good intra-rater reliability and inter-rater reliability for assessment of the muscle echogenicity of the supraspinatus and infraspinatus muscles. The correlation coefficients between B-mode ultrasound findings and MRI findings showed medium to large effect sizes (r=0.4-0.8). The Goutallier classification of the infraspinatus muscles was the MRI predictor with the best discriminative power for surgical reparability (area under the curve [AUC], 0.89; 95% confidence interval [CI], 0.81 to 0.98), while the Heckmatt scale for infraspinatus muscles was the most accurate ultrasound predictor (AUC, 0.85; 95% CI, 0.74 to 0.96). No significant differences in AUCs among the MRI and ultrasound predictors were found. Conclusion: B-mode ultrasonography was a reliable examination tool and had a similar ability to predict surgical reparability to that of MRI among patients with large-to-massive RCTs. Purpose: This study aimed to compare the ability of B-mode ultrasonography and magnetic resonance imaging (MRI) to predict the repairability of large-to-massive rotator cuff tears (RCTs).Methods: This cross-sectional study included participants with large-to-massive RCTs who underwent arthroscopic repair. B-mode ultrasonography and MRI were conducted prior to arthroscopic repair. B-mode ultrasonography was used to evaluate the echogenicity of the rotator cuff muscle using the Heckmatt scale. Intra-rater and inter-rater reliabilities were examined for two independent physicians. MRI was used to evaluate the degrees of tendon retraction, fatty infiltration of rotator cuff muscles, and muscle atrophy. Finally, two experienced orthopedic surgeons performed surgery and decided whether the torn stump could be completely repaired intraoperatively.Results: Fifty participants were included, and 32 complete repairs and 18 partial repairs were performed. B-mode ultrasonography showed good intra-rater reliability and inter-rater reliability for assessment of the muscle echogenicity of the supraspinatus and infraspinatus muscles. The correlation coefficients between B-mode ultrasound findings and MRI findings showed medium to large effect sizes (r=0.4-0.8). The Goutallier classification of the infraspinatus muscles was the MRI predictor with the best discriminative power for surgical reparability (area under the curve [AUC], 0.89; 95% confidence interval [CI], 0.81 to 0.98), while the Heckmatt scale for infraspinatus muscles was the most accurate ultrasound predictor (AUC, 0.85; 95% CI, 0.74 to 0.96). No significant differences in AUCs among the MRI and ultrasound predictors were found.Conclusion: B-mode ultrasonography was a reliable examination tool and had a similar ability to predict surgical reparability to that of MRI among patients with large-to-massive RCTs.

Lentivirus-mediated microRNA-124 gene-modified bone marrow mesenchymal stem cell transplantation promotes the repair of spinal cord injury in rats

Jia-Lin Song,Wei Zheng,Wei Chen,Yun Qian,Yuan-Ming Ouyang,Cun-Yi Fan 생화학분자생물학회 2017 Experimental and molecular medicine Vol.49 No.-

Our study aims to explore the effects of lentivirus-mediated microRNA-124 (miR-124) gene-modified bone marrow mesenchymal stem cell (BMSC) transplantation on the repair of spinal cord injury (SCI) in rats. BMSCs were isolated from the bone marrow of rats. The target gene miR-124 was identified using a luciferase-reporter gene assay. Seventy-two rats were selected for construction of the SCI model, and the rats were randomly divided into the blank group, sham group, SCI group, negative control (NC) group, overexpressed miR-124 group and si-PDXK group. The mRNA expression of miR-124 and the mRNA and protein expression of pyridoxal kinase (PDXK) were detected by quantitative real-time polymerase chain reaction and western blotting. The locomotor capacity of the rats was evaluated using the Basso, Beattie and Bresnahan (BBB) scale. Brdu, neuron-specific enolase (NSE), neurofilament (NF) and microtubule-associated protein 2 (MAP2) were detected using immunohistochemistry. The expression levels of thyrotropin-releasing hormone (TRH), prostacyclin (PGI2) and gangliosides (GM) were measured using an enzyme-linked immunosorbent assay. PDXK was identified as the target gene of miR-124. The overexpressed miR-124 group exhibited higher miR-124 expression than the SCI, NC and si-PDXK groups. Compared with the SCI and NC groups, the PDXK expression was downregulated in the overexpressed miR-124 and si-PDXK groups, and the BBB scores were significantly increased 7, 21 and 35 days after transplantation. The double-labeled positive cell densities (Brdu+NSE/NF/MAP2) and the expression levels of TRH, PGI2 and GM in the overexpressed miR-124 group were significantly higher than those in the NC and SCI groups. These results indicated that miR-124 targeted PDXK to accelerate the differentiation of BMSCs into neurocytes and promote SCI repair.

Comparison of Apixaban and Low Molecular Weight Heparin in Preventing Deep Venous Thrombosis after Total Knee Arthroplasty in Older Adults

Ting Guo,Jian-ning Zhao,Hui Jiang,Jia Meng,Yi-cun Wang,Jun Wang,Yang Qiu,Hao Ding 연세대학교의과대학 2019 Yonsei medical journal Vol.60 No.7

Purpose: To compare the effect of apixaban and low molecular weight heparin (LMWH) in the prevention and treatment of deepvenous thrombosis (DVT) after total knee arthroplasty in older adult patients. Materials and Methods: A total of 220 patients (average age of 67.8±6.4 years) undergoing total knee arthroplasty were randomlyselected as research subjects and were divided into apixaban and LMWH groups (110 in each group). Results: The incidence of DVT was lower in the apixaban group than in the LMWH group (5.5% vs. 20.0%, p=0.001). Activatedpartial thromboplastin times (35.2±3.6 sec vs. 33.7±2.2 sec, p=0.010; 37.8±4.6 sec vs. 34.1±3.2 sec, p<0.001; 39.6±5.1 sec vs. 35.7±3.0sec, p=0.032) and prothrombin times (14.0±1.0 sec vs. 12.8±0.9 sec, p<0.001; 14.5±1.2 sec vs. 13.0±1.1 sec, p<0.001; 15.3±1.4 sec vs. 13.2±1.3 sec, p=0.009) in the apixaban group at 1 week after surgery, 3 weeks after surgery, and the end of treatment were higherthan those in the LMWH group. Platelet and fibrinogen levels in the apixaban group were lower than those of the LMWH group. Also, capillary plasma viscosity and erythrocyte aggregation in the apixaban group at 1 week after surgery, 3 weeks after surgery,and the end of treatment were lower than those in the LMWH group. Conclusion: Apixaban, which elicits fewer adverse reactions and is safer than LMWH, exhibited better effects in the preventionand treatment of DVT after total knee arthroplasty in older adults.

Serum Peroxiredoxin3 is a Useful Biomarker for Early Diagnosis and Assessemnt of Prognosis of Hepatocellular Carcinoma in Chinese Patients

Shi, Liang,Wu, Li-Li,Yang, Jian-Rong,Chen, Xiao-Fei,Zhang, Yi,Chen, Zeng-Qiang,Liu, Cun-Li,Chi, Sheng-Ying,Zheng, Jia-Ying,Huang, Hai-Xia,Yu, Fu-Jun,Lin, Xiang-Yang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.7

Background: Recently, peroxiredoxin3 (PRDX3) was identified as a novel molecular marker for the progression of hepatocellular carcinoma (HCC). However, its potential clinical application as a serum marker for the early diagnosis and prognosis of HCC has not been investigated. Methods: PRDX3, alpha-fetaprotein (AFP), and other biochemical parameters were measured in serum samples from 297 Chinese patients, including 96 with HCC, 98 with liver cirrhosis (LC), and 103 healthy controls (HCs). Correlations between serum PRDX3 expression and clinicopathological variables and the relationship between serum PRDX3 expression and prognosis were analyzed. Results: Serum PRDX3 was significantly higher in HCC patients than in the LC and HC groups. The sensitivity and specificity of serum PRDX3 for the diagnosis of HCC were 85.9% and 75.3%, respectively, at a cutoff of 153.26 ng/mL, and the area under the curve was 0.865. Moreover, serum PRDX3 expression was strongly associated with AFP level, tumor diameter, TNM stage, and portal vein invasion. Kaplan-Meier curve analysis revealed that HCC patients with high serum PRDX3 expression had a shorter median survival time than those with low PRDX3 expression. Moreover, serum PRDX3 expression was an independent risk factor for overall survival. The inverse correlation between serum PRDX3 and patient survival remained significant in patients with early-stage HCC and in those with normal serum AFP levels. Conclusions: Serum PRDX3 can be used as a noninvasive biomarker for the diagnosis and/or prognosis of HCC.

Influence of Curcumin on HOTAIR-Mediated Migration of Human Renal Cell Carcinoma Cells

Pei, Chang-Song,Wu, Hong-Yan,Fan, Fan-Tian,Wu, Yi,Shen, Cun-Si,Pan, Li-Qun Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.10

Background: This study investigated the influence of curcumin on HOX transcript antisense RNA (HOTAIR)-mediated migration of cultured renal cell carcinoma (RCC) cells. Materials and Methods: Five RCC cell lines (769-P, 769-P-vector, 769-P-HOTAIR, 786-0, and Kert-3 ) were maintained in vitro. The expression of HOTAIR mRNA was determined by quantitative real-time PCR and cell migration was measured by transwell migration assay. The effects of different concentrations of curcumin (0 to $80{\mu}mol/L$) on cell proliferation was determined by the CCK-8 assay and influence of non-toxic levels (0 to $10{\mu}M$) on the migration of RCC cells was also determined. Results: Comparison of the 5 cell lines indicated a correlation between HOTAIR mRNA expression and cell migration. In particular, the migration of 769-P-HOTAIR cells was significantly higher than that of 769-P-vector cells. Curcumin at $2.5-10{\mu}M$ had no evident toxicity against RCC cells, but inhibited cell migration in a concentration-dependent manner. Conclusions: HOTAIR expression is correlated with the migration of RCC cells, and HOTAIR may be involved in the curcumin-induced inhibition of RCC metastasis.