1.Effect of palmitate on TXNIP expression in INS-1 islet cells
Qian ZHANG ; Nannan LIANG ; Xiangzheng WU ; Jin WANG ; Jiahui ZHAO ; Xiangying JIAO
Chinese Journal of Pathophysiology 2017;33(5):908-912
AIM:Chronic exposure to elevated levels of free fatty acids (FFAs) in type 2 diabetes patients is toxic to pancreatic β-cells.Thioredoxin (Trx)-interacting protein (TXNIP), an endogenous Trx-inhibiting protein, is up-regulated by glucose and is a critical mediator of hyperglycemia-induced β-cell apoptosis in diabetes.However, the effects of FFAs on TXNIP are unknown.In this experiment we observed the effect of palmitate on TXNIP expression in cultured INS-1 islet cells and the pathways involved were analyzed meanwhile.METHODS:After the full basis of preliminary experiment of incubating INS-1 cells with palmitate at different concentrations for different time, INS-1 islet cells were cultured with 0.5 mmol/L palmitate for 24 h.TXNIP expression, cell apoptosis, and expression of transcription factors related to TXNIP transcriptional regulation were determined.RESULTS:Compared with control group, the expression of TXNIP at mRNA and protein levels in palmitate group was significantly up-regulated (P<0.01).Cleaved caspase-3/caspase-3 ratio was increased in palmitate group (P<0.05), and the apoptosis of the INS-1 cells was also significantly increased (P<0.01).Palmitate enhanced the phosphorylation of nuclear factor-κB (NF-κB) (P<0.01), and the NF-κB inhibitors, PDTC and SN50, both blocked the palmitate-induced up-regulation of TXNIP expression.CONCLUSION:Saturated fatty acid palmitate enhances the expression of TXNIP.The mechanism of palmitate-induced TXNIP expression may be associa-ted with the increase in NF-κB phosphorylation.
2.Protective effect of hydrogen gas on neurons in rat hippocampus CA1 region during global cerebral ischemia/reperfusion injury
Nannan YUAN ; Yuning XIA ; Xinlei ZHANG ; Wei LIANG ; Youzhen WEI ; Yongxing TAN
The Journal of Practical Medicine 2016;32(6):870-874
Objective To investigate the effect of high concentration hydrogen gas on neurons in the rat hippocampus CA1 region during global cerebral ischemic/reperfusion injury (GCIR) Methods Four-vessel occlusion was used to establish rat model with GCIR injury. One hundred and five healthy male Sprague-Dawley rats were randomly divided into sham operation group(SH group, n = 15), model group(4-VO group, n = 45) and treatment group(4-VO+H2 group,n = 45). After 72 h and 9 d reperfusion, hippocampal CA1 region pyramidal neurons in every group were detected with Nissle staining , immunohistochemical neuron-specific nuclear protein (NeuN), specific protein antibody microglial cells (Iba1) staining and the relationship of position between neurons and microglia was observed through fluorescence double staining. We used Morris water maze to test the space orientation ability and the learning and memory ability in rats after 9 d reperfusion. Results Compared with those of 4-VO group,the neurons of hippocampus CA1 region were closer to normal in 72 h and 9 d in 4-VO+H2 group and neuron form and the number of neuron survival were increased significantly (P < 0.05);immunohistochemical staining showed that the number of neuron survival in 4-VO+H 2 group was obviously higher than that in 4-VO group (P < 0.05) and the number of microglia in 4-VO group was obviously higher than that in 4-VO+H2 group (P < 0.05). Water maze experiment showed that the swimming time in quadrant Ⅳ in 4-VO+H2 group was longer than that in 4-VO group (P < 0.05). Conclusion Inhalation of high concentration hydrogen gas has prominent protective effect on neurons of rat hippocampal CA1 region during reperfusion. The mechanism may be related with inhibiting the microglia excitation and activation during GCIR.
3.Mechanisms of erythropoietin-producing hepatocellular A3 participating in the invasion of hepatocellular carcinoma cells via regulating vascular endothelial growth factor
Liang ZHOU ; Desheng WANG ; Hui ZHAO ; Nannan HE ; Mingwen KOU ; Kefeng DOU
Chinese Journal of Digestive Surgery 2014;13(3):207-212
Objective To investigate the mechanisms of erythropoietin-producing hepatocellular A3 (EphA3) in the invasion of hepatocellular carcinoma (HCC) cells.Methods Hepatic cell HL-7702 and HCC cell and HCC cell lines HepG2 and MHCC97H were cultured.The expression of EphA3 in the HepG2 and MHCC97H cells was suppressed by siRNA interference,and then were divided into the untreated group,the control group and the siRNA intervention group.The expression of EphA3 was detected by RT-PCR and Western blot.The invasion ability of HepG2 and MHCC97H was detected by Transwell chamber.The protein expression of VEGF and activity of vascular endothelial growth factor (VEGF) were detected by western blot and ELISA.All data were analyzed using the analysis of variance or LSD-t test.Results The relative mRNA expressions of EphA3 in HL-7702,HepG2,and MHCC97H cells were 0.94 ±0.13,1.76 ±0.16 and 3.62 ±0.14,respectively,and the protein expressions of EphA3 in the 3 cells were 0.96 ±0.12,1.59 ±0.11 and 3.82 ±0.11.There was significant difference in the EphA3 expression between HL-7702 cells and HepG2,MHCC97H cells (t =2.511,6.437 ; 2.321,6.895,P < 0.05).The relative mRNA expressions of EphA3 in the HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.95 ±0.11,0.96 ±0.12 and 0.31 ±0.15,respectively.There was significant difference in the mRNA expression of EphA3 in the HepG2 cells between the siRNA intervention group and the control group (t =4.051,P < 0.05).The relative mRNA expressions of EphA3 in the MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.97 ± 0.16,0.95 ± 0.14 and 0.40 ± 0.11,respectively.There was significant difference in the mRNA expression of EphA3 in the MHCC97H cells between the siRNA interference group and the control group (t =5.237,P <0.05).The relative protein expressions of EphA3 in the HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.97 ± 0.16,0.95 ± 0.15 and 0.32 ± 0.17,respectively.There was significant difference in the protein expression of EphA3 in the HepG2 cells between the siRNA interference group and the control group (t =4.145,P < 0.05).The relative protein expressions of EphA3 in the MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.95 ± 0.11,0.96 ± 0.12 and 0.38 ±0.17,respectively.There was significant difference in the protein expressions of EphA3 in the MHCC97H cells between the siRNA interference group and the control group (t =4.327,P < 0.05).The numbers of HepG2 cells penetrated the Watrigel in the untreated group,the control group and the siRNA intervention group were (111 ±4)/10HPF,(109 ±5)/10HPF and (51 ±3)/10HPF,respectively.There was significant difference in the number of HepG2 cells between the siRNA interference group and the control group (t =7.582,P < 0.05).The numbers of MHCC97H cells penetrated the Watrigel in the untreated group,the control group and the siRNA intervention group were (402 ± 6)/10HPF,(397 ± 7)/10HPF and (152 ± 7)/10HPF,respectively.There was significant difference in the number of MHCC97H cells between the siRNA interference group and the control group (t =9.479,P < 0.05).The relative protein expressions of VEGF in the HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.98 ± 0.11,0.96 ± 0.13 and 0.57 ± 0.11,respectively.There was significant difference in the protein expression of VEGF of the HepG2 cells between the siRNA interference group and the control group (t =3.167,P < 0.05).The relative protein expression of VEGF in the MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.97 ±0.14,0.98 ±0.12 and 0.34 ± 0.15,respectively.There was significant difference in the protein expression of VEGF of the MHCC97H cells between the siRNA interference group and the control group (t =4.278,P < 0.05).The relative activities of VEGF proteins of HepG2 cells in the untreated group,the control group and the siRNA intervention group were 0.96 ±0.15,0.94 ±0.11 and 0.47 ±0.13,respectively.There was significant difference in the activity of VEGF protein in the HepG2 cells between the siRNA interference group and the control group (t =3.981,P < 0.05).The relative activities of VEGF proteins in MHCC97H cells in the untreated group,the control group and the siRNA intervention group were 0.98 ±0.12,0.97 ±0.12 and 0.38 ±0.14,respectively.There was significant difference in the activity of VEGF protein in the MHCC97H cells between the siRNA interference group and the control group (t =4.059,P < 0.05).Conclusions EphA3 plays an important role in the invasion of HCC cells via regulating the protein expression and activity of VEGF.EphA3 might be a new target for the treatment of HCC.
4.A trail of transcatheter closure of left ventricular-right atrium communication
Nannan MU ; Yuanyuan SUN ; Xiaozhou ZHENG ; Bo ZHANG ; Jiali LIANG ; Haofeng SUN
Chinese Journal of Ultrasonography 2013;22(7):580-582
Objective To review and summarize the experiences of transcatheter closure of left ventricular-right atrium communication,and discuss the feasibility of interventional therapy for this kind of cardiac abnormalitis.Methods 22 patients who suffered from left ventricular-right atrium communication underwent transcatheter interventional therapy with ventricular septal defect(VSD) occluder.The operating procedures were performed as like as the transcatheter closure of VSD:established the pathway from femoral artery to femoral vein through left ventricle,VSD,right atrium (or from left ventricle to right atrial through the communication) and inferior vena cava,then inserted the polysheath from femoral vein,introduced by the pathway to left ventricle,and implanted VSD occluder through the polysheath to close the shunt.Results The operation succeeded in 21 patients.The cardiac murmur was disappeared in all patients,and there was no residual shunt or aortic regurgitation that conformed by postoperative ventriculography and echocardiography in follow up phase,and the tricuspid regurgitation was lessened than preoperative.The operation abort in 1 patient because of aortic regurgitation after implanting occluder.Conclusions Transcatheter closure of left ventricular-right atrium communication is feasible as the selected occluder is accordant,and atrioventricular block,aortic regurgitation and tricuspid regurgitation can be avoided.
5.The surface structure and antibacterial property of Ag-containing coating on casting cobalt chromium alloy
Ruiying LIANG ; Jingdong LI ; Yanping ZHAO ; Yanli XU ; Wenhui WU ; He MENG ; Nannan LIAO
Journal of Practical Stomatology 2014;(5):637-640
Objective:To study the surface structure and antibacterial property of Ag-containing coating(AgCC)on casting cobalt chromium alloy(Co-Cr).Methods:Plasma spraying technique was adopted to prepare AgCC on Co-Cr.Scanning electron microsco-py,energy dispersive analysis and X-ray diffraction analysis were used to evaluate the surface properties.The antibacterial effects of AgCC against staphylococcus aureus and candida albicans were examined by film attachment method.Fluorescence microscope was a-dopted to evaluate the bacteria adhesion property.Results:AgCC surface was uniform and compact,combined perfectly with sub-strate material.The content of the surface was mainly Ag,Cr and a small amount of Ag2 O and Cr2 O3 .The results of antibacterial as-say showed that on AgCC the number of bacterial colony was fewer(P<0.05)and bacterial adhesion was less than those on Co-Cr(P<0.05).Conclusion:AgCC surface was stable and has antibacterial property.
6.Time-dependent changes in CT of radiation-induced liver injury: A preliminary study in gastric cancer patients.
Yaqi, SHENG ; Qiuxia, WANG ; Zhen, LI ; Nannan, ZHENG ; Yigang, PEI ; Liang, CHEN ; Dayu, HU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):683-6
In this study, the time-dependent changes on dynamic computed tomograph (CT) of radiation-induced liver injury in gastric cancer patients was examined. The CT images of 52 gastric cancer patients who had received chemoradiotherapies were reviewed on the PACS system. Dynamic CT scan was performed in all the subjects. Our results showed that 18 patients were found to have radiation-induced liver injury. The CT findings of radiation-induced liver injury in gastric cancer patients tend to show up one month after radiation treatment. The damaged area was of low density on all three phases, and then it was enhanced on portal vein phase or delay phase. The focal radiation reaction of liver without basic disease vanished 9-11 months later after treatment. We are led to conclude that dynamic CT is of help in the diagnosis of CRT-induced liver injury, and it may be the method of choice for following up the whole course of the CRT-induced liver injury, i.e., form hepatic damage to healing. The classification of CT findings we recommend can avoid the influence of technological factors, and thereby serve as a better guide for treatment of CRT-induced liver injury.
7.Panax japlcus var polysaccharide regulates proliferation and apoptosis of gastric cancer MKN45 cells by targeting let-7a/CDK6 molecular axis
WANG Bingshu ; LIANG Rongzhen ; JI Nannan
Chinese Journal of Cancer Biotherapy 2020;27(2):135-141
Objective: To investigate the effect of panax japlcus var polysaccharide (PJPS) on the proliferation and apoptosis of gastric cancer MKN45 cells and its regulatory mechanism. Methods: Human gastric cancer cell lines (HGC27, MGC803, MKN45) and gastric mucosal epithelial cell line GES-1 were selected for this study. Let-7a mimics and let-7a inhibitor were transfected into MKN45 cells; Gastric cancer cell lines were treated with 100 μg/ml PJPS and MKN45 was selected as the subsequent experimental cell line. MKN45 cells were cultured with 0, 10, 50, 100 and 120 μg/ml PJPS, respectively. The proliferation and apoptosis rate of MKN45 cells were detected by CCK-8 and flow cytometry, respectively. Expressions of cell cycle dependent kinase 6 (CDK6) and apoptosis-related proteins in MKN45 cells were detected by Western blotting, and the expression level of miRNAs regulating the proliferation of gastric cancer cells was detectedbyReal-timequantitativePCR(qPCR).TheDualluciferasereportergeneassaywasusedtovalidatethetargeting relationship between let-7a and CDK6. Results: Compared with other gastric cancer cells, 100 μg/ml PJPS significantly inhibited the proliferation of MKN45 cells (P<0.01). At the same time, 100 μg/ml PJPS significantly up-regulated the expression of let-7a in MKN45 cells (P<0.01). The Dual luciferase reporter gene assay confirmed that CDK6 was the target gene of let-7a. Furthermore, PJPS inhibited the expression of CDK6 by up-regulating let-7a, thereby inhibiting the proliferation and inducing apoptosis of MKN45 cells (all P<0.01). Conclusion: PJPS inhibits proliferation and induces apoptosis of gastric cancer MKN45 cells by regulating the let-7a/ CDK6 axis.
8.Time-dependent Changes in CT of Radiation-induced Liver Injury: A Preliminary Study in Gastric Cancer Patients
SHENG YAQI ; WANG QIUXIA ; LI ZHEN ; ZHENG NANNAN ; PEI YIGANG ; CHEN LIANG ; HU DAYU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):683-686
In this study, the time-dependent changes on dynamic computed tomograph (CT) of radiation-induced liver injury in gastric cancer patients was examined. The CT images of 52 gastric cancer patients who had received chemoradiotherapies were reviewed on the PACS system. Dynamic CT scan was performed in all the subjects. Our results showed that 18 patients were found to have radiation-induced liver injury. The CT findings of radiation-induced liver injury in gastric cancer patients tend to show up one month after radiation treatment. The damaged area was of low density on all three phases, and then it was enhanced on portal vein phase or delay phase. The focal radiation reaction of liver without basic disease vanished 9-11 months later after treatment. We are led to conclude that dynamic CT is of help in the diagnosis of CRT-induced liver injury, and it may be the method of choice for following up the whole course of the CRT-induced liver injury, i.e., form hepatic damage to healing. The classification of CT findings we recommend can avoid the influence of technological factors, and thereby serve as a better guide for treatment of CRT-induced liver injury.
9.Retroperitoneoscopic dismembered pyeloplasty for pediatric ureteropelvic junction obstruction: a report of 85 cases.
Zhi CHEN ; Xiang CHEN ; Lin QI ; Liang CHEN ; Yancheng LUO ; Yao HE ; Nannan LI ; Chaoqun XIE
Journal of Central South University(Medical Sciences) 2011;36(5):430-434
OBJECTIVE:
To investigate the clinical application of retroperitoneoscopic dismembered pyeloplasty for pediatric ureteropelvic junction obstruction (UPJO).
METHODS:
Of the 85 pediatric patients with UPJO, 56 were boys and 29 were girls. The age of the patients ranged from 2.5 to 11 years (mean = 4.6 years).B-ultrasonography showed hydronephrosis <15 mm (mild) in 15 patients,15-30 mm (moderate) in 59, <30 mm (severe) in the other 6. Intravenous urography (IVU) showed good imaging in 55 patients within 30 min, light imaging in 27 at 30-120 min, and no imaging in 3 after 120 min. The 85 patients underwent retroperitoneoscopic dismembered pyeloplasty. Conventional antegrade or modified antegrade double-J stenting was inserted in the 85 patients intraoperatively.
RESULTS:
The operation was successful in all, with no conversion to open surgery during the operation. The mean operation time was 146 min (125-240 min).The mean blood loss was 68 mL (55-112 mL).The mean postoperative hospitalization was 7 d (6-8 d ).Urine leakage occurred in 1 patient and with a good drainage, urine leakage disappeared in 1 week. Follow-up ranged 3-24 months (mean = 11 months).IVU showed on UPJ stricture, and good imaging in 75 patients within 30 min, light imaging in 10 at 30-120 min. Hydronephrosis was remitted:hydronephrosis resolution in 51 patients, mild in 6 and moderate in 3.
CONCLUSION
Retroperitoneoscopic dismembered pyeloplasty is a safe, effective and mini-invasive procedure for pediatric ureteropelvic junction obstruction, with a rapid postoperative recovery. It will be the ideal treatment for UPJO in pediatric patients.
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10.Protective effect of hydrogen gas on neurons and dendritic spines of hippocampus CA1 region in rats after global cerebral ischemia/reperfusion injury
Yongxing TAN ; Nannan YUAN ; Yuning XIA ; Xinlei ZHANG ; Wei LIANG ; Youzhen WEI
Chinese Journal of Neuromedicine 2016;15(11):1091-1097
Objective To explore the protective effect of in-taking high concentration hydrogen gas on neurons and dendritic spines in hippocampus CA1 region of rats after globe cerebral ischemia/reperfusion (I/R) injury and its mechanism.Methods Four-vessel occlusion (4VO) was used to establish the models of global cerebral I/R injury in rats.One hundred and twenty healthy male Sprague-Dawley rats were randomly divided into 3 groups using a random number table:sham-operated group (inhaled 67% N2 and 67% O2,n=40),model group (inhaled 67% N2 and 67% O2 during reperfusion,n=40),and treatment group (inhaled 67% H2 and 67% O2 during reperfusion,n=40).After 72 h,5 and 9 d reperfusion,neuron-specific nuclear (NeuN) protein expression in the pyramidal neurons of the hippocampal CA1 region was detected with immumohistochemical staining and the positive cells were counted.And the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum were tested with colorimetry.Water maze test was used to measure the spatial orientation and memory function and Golgi staining to detect the number of dendritic spines in neurons 9 d after reperfusion.Results (1) Immunohistochemical staining of NeuN results showed that as compared with those in the model group,the neurons ofhippocampus CA1 region were significantly closer to normal with relatively intact structure,and the number of positive neurons was significantly increased in the treatment group 72 h,5 d,and 9 d after reperfusion (P<0.05).With the reperfusion time being prolonged,the number of NeuN stained positive neurons at different time points of reperfusion in model group was gradually decreased (P<0.05),and the numeric of the NeuN stained positive neurons at different time points of reperfusion in treatment group was slightly declined without significant difference (P>0.05).(2) The serum SOD activity in the treatment group was significantly higher than that in the model group and sham-operated group (P<0.05),while the MDA content in the treatment group was significantly lower than that in model group 72 h,5 d,and 9 d after reperfusion (P<0.05).And with the reperfusion time being prolonged,the SOD activity at different time points of reperfusion showed no significant difference among the difference groups (P>0.05).But the MDA content at different time points ofreperfusion between model group and treatment group was significantly different (P<0.05);with the reperfusion time being prolonged,the MDA content was gradually decreased in both groups.(3) Nine d after reperfusion,water maze test found that the incubation period of treatment group was significantly shorter than that of model group (P<0.05);the IV quadrant swimming time of space exploration in the treatment group was significantly longer than that in the model group (P<0.05).(4) Golgi staining showed that the complexity of the neuronal dendrites branch and the number of dendritic spines of neurons in the hippocampal CA1 region of treatment group were increased than those in the model group;high-power oil microscopy indicated that the density of dendritic spines in the treatment group was significantly higher than that in the model group (P<0.05).Conclusion In-taking of high concentrations hydrogen gas during reperfusion can definitely protect neurons in hippocampal CA1 region after globe cerebral I/R injury,and improve learning and memory function,whose mechanism may be related to hydrogen protecting the structure and function of neurons and dendritic spines,and inhibiting oxidative stress to reduce oxidative damage.