查看更多>>摘要：Microtubule-associated serine/threonine kinase(MASTL)functions to regulate chromosome condensation and mitotic progression.Therefore,aberrant MASTL expression is commonly implicated in various human cancers.This study analyzed MASTL expression in gastric cancer vs.adjacent normal tissue for elucidating the association with clinicopathological data from patients.This work was then extended to investigate the effects of MASTL knockdown on tumor cells in vitro.The level of MASTL expression in gastric cancer tissue was assessed from the UALCAN,GEPIA,and Oncomine online databases.Lentivirus carrying MASTL or negative control shRNA was infected into gastric cancer cells.RT-qPCR,Western blotting,cell viability,cell counting,flow cytometric apoptosis and cell cycle,and colony formation assays were performed.MASTL was upregulated in gastric cancer tissue compared to the adjacent normal tissue,and the MASTL expression was associated with advanced tumor stage,Helicobacter pylori infection and histological subtypes.On the other hand,knockdown of MASTL expression significantly reduced tumor cell viability and proliferation,and arrested cell cycle at G2/M stage but promoted tumor cells to undergo apoptosis.At protein level,knockdown of MASTL expression enhanced levels of cleaved PARP1,cleaved caspase-3,Bax and p-ERKl/2 expression,but downregulated expression levels of BCL-2 and p-NF-κB-p65 protein in AGS and MGC-803 cells.MASTL overexpression in gastric cancer tissue may be associated with gastric cancer development and progression,whereas knockdown of MASTL expression reduces tumor cell proliferation and induces apoptosis.Further study will evaluate MASTL as a potential target of gastric cancer therapeutic strategy.
查看更多>>摘要：The incidence of colorectal cancer(CRC)is increasing in China,with high mortality.Here,we aimed to evaluate the latest clinicopathological features and prognostic value of the KRAS/NRAS/BRAF mutation status in CRC patients in Central China.The clinical data of 1549 CRC patients with stage Ⅰ-Ⅳ disease diagnosed at Union Hospital,Tongji Medical College of Huazhong University of Science and Technology from 2015 to 2017 were collected and analyzed retrospectively.KRAS/NRAS/BRAF mutations were detected by real-time quantitative polymerase chain reaction(q-PCR)in 410 CRC patients,with mutation frequencies of KRAS,NRAS and BRAF of 47.56％,2.93％and 4.15％,respectively.The gene mutation status and clinicopathological characteristics of 410 patients with CRC who underwent qPCR were analyzed.The KRAS and BRAF gene mutations were related to the pathological differentiation and number of metastatic lymph nodes.The BRAF gene mutation was also associated with cancer thrombosis in blood vessels.Cox regression analysis showed that there was no statistically significant difference in the overall survival(OS)between patients with KRAS,NRAS mutants and wild-type CRC patients,while the BRAF gene mutation was negatively correlated with the OS rate of CRC patients.It is suggested that the BRAF gene mutation may be an independent risk factor for the prognosis of CRC.
查看更多>>摘要：The incidence of colorectal cancer(CRC)is increasing in China,with high mortality.Here,we aimed to evaluate the latest clinicopathological features and prognostic value of the KRAS/NRAS/BRAF mutation status in CRC patients in Central China.The clinical data of 1549 CRC patients with stage Ⅰ-Ⅳ disease diagnosed at Union Hospital,Tongji Medical College of Huazhong University of Science and Technology from 2015 to 2017 were collected and analyzed retrospectively.KRAS/NRAS/BRAF mutations were detected by real-time quantitative polymerase chain reaction(q-PCR)in 410 CRC patients,with mutation frequencies of KRAS,NRAS and BRAF of 47.56％,2.93％and 4.15％,respectively.The gene mutation status and clinicopathological characteristics of 410 patients with CRC who underwent qPCR were analyzed.The KRAS and BRAF gene mutations were related to the pathological differentiation and number of metastatic lymph nodes.The BRAF gene mutation was also associated with cancer thrombosis in blood vessels.Cox regression analysis showed that there was no statistically significant difference in the overall survival(OS)between patients with KRAS,NRAS mutants and wild-type CRC patients,while the BRAF gene mutation was negatively correlated with the OS rate of CRC patients.It is suggested that the BRAF gene mutation may be an independent risk factor for the prognosis of CRC.
查看更多>>摘要：The stemness of different side population(SP)cell subtypes in ovarian cancer cells was studied,and the heterogeneity of ovarian cancer stem cells was analyzed.The cisplatin-resistant human serous ovarian cancer cell line C13 was stained with the bisbenzimide Hoechst 33342.A flow cytometry-based fluorescence-activated sorting method was used to obtain lower-SP(LSP)cells,upper-SP(USP)cells,and non-SP cells(NSP)based on their sensitivity to the staining time and Hoechst dye concentration.The sphere-forming capability,expression levels of stem cell markers,resistance to high concentrations of cisplatin,and subcutaneous tumorigenicity in NOD/SCID mice of the different cell subtypes were evaluated.The C13 cells contained SP cells with stemness characteristics,and the LSP cell subtype expressed higher levels of stem cell markers,had higher in vitro sphere-forming capability,higher cisplatin resistance and higher in vivo subcutaneous tumorigenesis than USP cells(P＜0.05).NSP cells had no stemness.In conclusion,different subtypes of ovarian cancer SP cells have different stemness levels,and ovarian cancer stem cells may be heterogeneous.
查看更多>>摘要：The stemness of different side population(SP)cell subtypes in ovarian cancer cells was studied,and the heterogeneity of ovarian cancer stem cells was analyzed.The cisplatin-resistant human serous ovarian cancer cell line C13 was stained with the bisbenzimide Hoechst 33342.A flow cytometry-based fluorescence-activated sorting method was used to obtain lower-SP(LSP)cells,upper-SP(USP)cells,and non-SP cells(NSP)based on their sensitivity to the staining time and Hoechst dye concentration.The sphere-forming capability,expression levels of stem cell markers,resistance to high concentrations of cisplatin,and subcutaneous tumorigenicity in NOD/SCID mice of the different cell subtypes were evaluated.The C13 cells contained SP cells with stemness characteristics,and the LSP cell subtype expressed higher levels of stem cell markers,had higher in vitro sphere-forming capability,higher cisplatin resistance and higher in vivo subcutaneous tumorigenesis than USP cells(P＜0.05).NSP cells had no stemness.In conclusion,different subtypes of ovarian cancer SP cells have different stemness levels,and ovarian cancer stem cells may be heterogeneous.
查看更多>>摘要：Triptolide,a component of the Chinese herb Tripterygium wilfordii Hook F,has been proved to be effective in the treatment of rheumatoid arthritis(RA).However,its underlying mechanisms on RA have not yet been well established.We observed the inhibitory effect of triptolide on the expression of inflammatory cytokines and proliferation of fibroblast-like synoviocytes(FLS)induced by the complex of interleukin-6(IL-6)and the soluble form of the IL-6 receptor(sIL-6R).Furthermore,to clarify the underlying mechanisms,we treated FLS with the Janus-activated kinase 2(JAK2)inhibitor/signal transducer and activator of transcription 3(STAT3)activation blocker AZD1480.In this study,immunohistochemical staining was used to identify vimentin(+)and CD68(-)in FLS.The FLS proliferation was measured by cell proliferation assay,and the cell cycles were analyzed by flow cytometry.Furthermore,ELISA was used to detect the expression of the inflammatory factors in culture solution.The expression levels of p-JAK2,JAK2,p-STAT3 and STAT3 were investigated through Western blotting analysis.The results showed that IL-6/sIL-6R significantly increased the cell proliferation and expression of inflammatory cytokines,including IL-6,interleukin-1β(IL-1β)and vascular endothelial growth factor(VEGF).Triptolide or AZD1480 inhibited the cell proliferation and inflammatory cytokine expression in IL-6/sIL-6R-stimulated FLS by suppressing JAK2/STAT3.The study suggested that the physiological effects of triptolide on RA were due to its contribution to the inhibition of the inflammatory cytokine expression and FLS proliferation by suppressing the JAK2/STAT3 signaling pathway.It may provide an innovative insight into the effect of triptolide in preventing RA pathogenesis.
查看更多>>摘要：Triptolide,a component of the Chinese herb Tripterygium wilfordii Hook F,has been proved to be effective in the treatment of rheumatoid arthritis(RA).However,its underlying mechanisms on RA have not yet been well established.We observed the inhibitory effect of triptolide on the expression of inflammatory cytokines and proliferation of fibroblast-like synoviocytes(FLS)induced by the complex of interleukin-6(IL-6)and the soluble form of the IL-6 receptor(sIL-6R).Furthermore,to clarify the underlying mechanisms,we treated FLS with the Janus-activated kinase 2(JAK2)inhibitor/signal transducer and activator of transcription 3(STAT3)activation blocker AZD1480.In this study,immunohistochemical staining was used to identify vimentin(+)and CD68(-)in FLS.The FLS proliferation was measured by cell proliferation assay,and the cell cycles were analyzed by flow cytometry.Furthermore,ELISA was used to detect the expression of the inflammatory factors in culture solution.The expression levels of p-JAK2,JAK2,p-STAT3 and STAT3 were investigated through Western blotting analysis.The results showed that IL-6/sIL-6R significantly increased the cell proliferation and expression of inflammatory cytokines,including IL-6,interleukin-1β(IL-1β)and vascular endothelial growth factor(VEGF).Triptolide or AZD1480 inhibited the cell proliferation and inflammatory cytokine expression in IL-6/sIL-6R-stimulated FLS by suppressing JAK2/STAT3.The study suggested that the physiological effects of triptolide on RA were due to its contribution to the inhibition of the inflammatory cytokine expression and FLS proliferation by suppressing the JAK2/STAT3 signaling pathway.It may provide an innovative insight into the effect of triptolide in preventing RA pathogenesis.
查看更多>>摘要：The effect of preoperative Double-J(DJ)ureteral stenting before flexible ureterorenoscopy(FURS)in the treatment for urinary stones was evaluated.We retrospectively enrolled 306 consecutive patients who underwent FURS from Jan.2014 to Dec.2017.All the patients were classified into two groups according to whether they had DJ ureteral stenting before FURS.Baseline characteristics(age,sex,stone location,stone size,surgical success rate,operation time,stone-free rate of the first day after surgery,stone-free rate of the first month after surgery,total complication rate)were compared using Chi-square test for categorical variables and Kruskal-Wallis test for continuous variables.In total,306 patients were included in this study.The group of DJ stenting before FURS included 203(66.3％)patients,and non-DJ stenting before FURS was observed in 103(33.7％)patients.The group of DJ stenting before FURS was significantly associated with a shorter operation time(53.8 vs.59.3 min,P＜0.001),a higher stone-free rate of the first day after surgery(69.0％vs.51.5％,P=0.003).However,statistical significant differences were not found in the age,sex,stone location,stone size,surgical success rate,stone-free rate of the first month after surgery(89.2％vs.81.6％,P=0.065)and total complication rate(5.4％vs.9.7％,P=0.161)between the two groups.Preoperative DJ ureteral stenting before FURS could reduce the operation time and increase stone-free rate of the first day after surgery.However,it might not benefit the stone-free rate of the first month after surgery and reduce the complication rate.Preoperative DJ stenting should be not routinely performed.
查看更多>>摘要：The effect of preoperative Double-J(DJ)ureteral stenting before flexible ureterorenoscopy(FURS)in the treatment for urinary stones was evaluated.We retrospectively enrolled 306 consecutive patients who underwent FURS from Jan.2014 to Dec.2017.All the patients were classified into two groups according to whether they had DJ ureteral stenting before FURS.Baseline characteristics(age,sex,stone location,stone size,surgical success rate,operation time,stone-free rate of the first day after surgery,stone-free rate of the first month after surgery,total complication rate)were compared using Chi-square test for categorical variables and Kruskal-Wallis test for continuous variables.In total,306 patients were included in this study.The group of DJ stenting before FURS included 203(66.3％)patients,and non-DJ stenting before FURS was observed in 103(33.7％)patients.The group of DJ stenting before FURS was significantly associated with a shorter operation time(53.8 vs.59.3 min,P＜0.001),a higher stone-free rate of the first day after surgery(69.0％vs.51.5％,P=0.003).However,statistical significant differences were not found in the age,sex,stone location,stone size,surgical success rate,stone-free rate of the first month after surgery(89.2％vs.81.6％,P=0.065)and total complication rate(5.4％vs.9.7％,P=0.161)between the two groups.Preoperative DJ ureteral stenting before FURS could reduce the operation time and increase stone-free rate of the first day after surgery.However,it might not benefit the stone-free rate of the first month after surgery and reduce the complication rate.Preoperative DJ stenting should be not routinely performed.
查看更多>>摘要：Diabetic retinopathy(DR)is a common cause of blindness all over the world.Bone marrow mesenchymal stem cells(BMSCs)have been considered as a promising strategy for retinal regeneration in the treatment of DR.However,the poor viability and low levels of BMSCs engraftment limit the therapeutic potential of BMSCs.The present study aimed to examine the direct induction of BMSCs differentiation into the cell types related to retinal regeneration by using soluble cytokine ciliary neurotrophic factor(CNTF).We observed remarkably increased expression of cellular retinaldehyde-binding protein(CRALBP)and retinoid isomerohydrolase(RPE65)in BMSCs treated with CNTF in vitro,indicating the directional differentiation of BMSCs into the retinal pigment epithelium(RPE)cells,which are crucial for retinal healing.In vivo,the diabetic rat model was established by use of streptozotocin(STZ),and animals treated with BMSCs+CNTF exhibited better viability and higher delivery efficiency of the transplanted cells than those treated with BMSCs injection alone.Similar to the in-vitro result,treatment with BMSCs and CNTF combined led to the differentiation of BMSCs into beneficial cells(RPE cells),and accelerated retinal healing characterized by the activation of rod photoreceptor cells and phagocytosis function of RPE cells.In conclusion,CNTF contributes to the differentiation of BMSCs into RPE cells,which may help overcome the current stem cell therapy limitations in the field of retinal regeneration.
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