Objective To investigate the protective effects of Apigenin (APG) on hepatic in diabetic rats.Methods The experimental diabetes model were made by intraperitoneal injecting STZ. The rats were randomly devided into six groups: a normal control group, a diabetes model control group, APG 5, 10, 20 and 40 mg/kg treated groups, and the drug was given by intraperitoneal injection. 4 weeks later, the body weight, ratio of hepatic weight and body weight were detected; the histopathological changes of hepatic tissue were observed by HE staining; the content of ALT/AST/LDH in serum were determined; the activity of SOD/GSH-Px/CAT and the content of MDA in hepatic tissue were determined.Results Compared with the diabetes model control group, the body weight of APG(10, 20, 40) mg/kg treated groups were significantly increased (261.3 ± 15.8 g, 274.2 ± 18.4 g, 265.9 ± 19.0 gvs. 250.8 ± 21.4 g,P<0.05 orP<0.01), and the ratio of hepatic weight and body weight were significantly decreased (27.7 ± 5.69, 26.2 ± 4.91, 27.3 ± 4.58vs. 32.9 ± 5.85,P<0.05 orP<0.01); the hepatic tissue histopathological changes of APG-treated groups were significantly improved; the level of ALT in serum of APG (5, 10, 20 and 40) mg/kg treated groups were significantly decreased (1 039.3 ± 453.9 U/L, 917.6 ± 445.2 U/L, 828.4 ± 309.5 U/L, 721.7 ± 318.3 U/Lvs. 1 205.2 ± 484.1 U/L;P<0.05 orP<0.01), AST were significantly decreased (97.8 ± 23.8 U/L, 90.1 ± 19.6 U/L, 81.7 ± 15.7 U/L, 86.4 ± 19.2 U/Lvs. 105.3 ± 25.7 U/L;P<0.05,P<0.01), LDH were also significantly decreased (983.7 ± 192.6 U/L, 918.3 ± 212.9 U/L, 830.4 ± 174.2 U/L, 871.8 ± 183.1 U/Lvs. 1 102.8 ± 211.6 U/L;P<0.05 orP<0.01); the activity of SOD in hepatic tissue of APG (10, 20, 40) mg/kg treated groups were significantly increased (10.5 ± 1.9 U/mg, 11.6 ± 2.1 U/mg, 10.5 ± 2.0 U/mgvs. 9.1 ± 1.8 U/mg,P<0.05,P<0.01), GSH-Px were significantly increased (14.2 ± 2.7 U/mg, 15.3 ± 2.9 U/mg, 14.6 ± 2.6 U/mgvs. 12.9 ± 2.3 U/mg;P<0.05, P<0.01), CAT were significantly increased (3.15 ± 0.90 U/mg, 3.58 ± 0.88 U/mg, 3.31 ± 1.09 U/mgvs. 2.58 ± 0.79 U/mg,P<0.05 orP<0.01), and the content of MDA was significantly decreased (5.03 ± 1.70 nmol/mg, 4.66 ± 1.51 nmol/mg, 4.73 ± 1.65 nmol/mg vs. 5.98 ± 1.62 nmol/mg;P<0.05 orP<0.01). And the treatment effect of APG 20 mg/kg groups was the most significant.Conclusion APG had antioxidant enzyme activity, besides its reducing the damage of free radical, and protecting the hepatic tissue of diabetic rats effects.

Objective: To explore the mechanism of Xuebijing injection in the treatment of acute paraquat poisoning by means of studying the expression of TNF-alpha, NF-kappa B, Caspase-3 and the changes of cell apoptosis rate detected by TUNEL in the lung tissue of acute paraquat-induced rats. Methods: On the base of random number table, 126 Wister rats weighing 220 g to 270 g were divided into 3 groups: (1) Control group: 42 rats, (2) Poisoned group: 42 rats, (3) Treatment group: 42 rats. On 1^st、3^rd、7^th、14^th、21^st、28^th、and 35^th day, six rats from each group were anaesthetized by intraperitoneal injection of chloral hydrate. To cut the chest and take the lung tissue samples. The expression levels of Tumor Necrosis Factor-alpha, Nuclear Factor-kappa B and Caspase-3 protein in lung tissue were detected by immunohistochemical staining, as well as apoptotic cell rate was detected by TUNEL staining. Results: The expression levels of Tumor Necrosis Factor-alpha, Nuclear Factor-kappa B, Caspase-3 protein and TUNEL staining in the lung tissue of the poisoned group was significantly higher than that of the control group (P<0.05) . Compared with the poisoned group, the expression of TNF-alpha, NF-kappa B, Caspase-3 and TUNEL in treatment group decreased significantly (P<0.05) , but they were still higher than those of the control group, and the difference was statistically significant compared with the control group (P<0.05) . Conclusion Apoptosis and TNF-alpha, NF-kappa B and Caspase-3 play an important role in lung injury of paraquat-induced rats. Xuebijing injection can inhibit the expression of TNF-alpha, NF-kappa B, Caspase-3 in lung tissue, reduce the apoptosis rate and alleviate the damage of lung tissue in paraquat-poisoning rats.

Objective To study the effects of heme oxygenase-1 transduced by cell penetrating peptide PEP-1 on renal injury in acute paraquat-induced rats.Methods The fusion protein PEP-1/HO-1 was manufactured by genetic engineering methods.One hundred and twenty-six healthy adult Sprague-Dawley (SD) rats (63 male,63 female) were raudomly divided into three groups:(1)Control group:42 rats,(2)Poisoned group:42 rats,(3)PEP-1/HO-1 intervention group:42 rats.The rats in Poisoned group and intervention group were treated intraperitoneally with paraquat (25 mg/kg) deliquated with normal saline.Control group rats were treated with the same way of normal saline as the others.Intervention group rats were injected 1 mg of the fusion protein PEP-1/HO-1 into the left iliac vein at 30 minutes before the administration of the paraquat.After abdominal dissection with ether anaesthesia,six rats respectively from each of three groups were taken tissue samples fron kidney at l st、6 th、12 th、24 th、36 th、48 th、and 72 nd hour respectively.Tissue Superoxide Dismutase (SOD) activity and Malondialdehyde (MDA) content were measured,and renal tissue was quickly remained to observe the expression of Heme Oxygenase-1 (HO-1) by the method of imnmnohistochenistry.Blood samples were collected from the abdominal aorta fur determination of Blood Urea Nitrogen (BUN) and Creatinine (Cr) concentrations in plasma.Results To prepare high purity fusion protein PEP-1/HO-1 Successfully.The levels of the plasma Bun and Cr in Poisoned and intervention group were significantly higher than that in Control group in the six to seventy-two hours (P＜0.01);Whereas the increases of Bun andi Cr were markedly intibited in PEP-1/HO-1 intervention group,in which,the levels of Bun and Cr in the six to seventytwo hours were significantly lower than Poisoned group (P＜0.05).Comnpared with control group,the levels of MDA in poisoned groups were increased in the one to forty-eight hours and the levels of it in groups PEP-1/HO-1 in the one to thirty-six(P＜0.01 or P＜0.05).The T-SOD activity of the poisoued group was decreased in the one to thirty-six hours than that of the control group (P＜0.05).Compared with poisoned group,the T-SOD activity at various points in time was increased in group PEP-1/HO-1 (P＜0.01).In control group,there was only very.weak expressions of HO-1 in the normal renal tissue.In poisoned and intervention group,the expressions of HO-1 were significantly higher than that in control group in all time (P＜0.01).The HO-1 expressions of PEP-1/HO-1 group in all time was significantly higher than that in poisoned group (P＜0.01 or P＜0.05).Conclusion The HO-1 protein Can be successfully transduced into renal tissue by cell penetrating peptide PEP-1 and the transduced HO-1 protein reduces renal injury of the acute paraquat-induced rats by inhibiting lipid peroxidation response.

Objective To study the effects of heme oxygenase-1 transduced by cell penetrating peptide PEP-1 on renal injury in acute paraquat-induced rats.Methods The fusion protein PEP-1/HO-1 was manufactured by genetic engineering methods.One hundred and twenty-six healthy adult Sprague-Dawley (SD) rats (63 male,63 female) were raudomly divided into three groups:(1)Control group:42 rats,(2)Poisoned group:42 rats,(3)PEP-1/HO-1 intervention group:42 rats.The rats in Poisoned group and intervention group were treated intraperitoneally with paraquat (25 mg/kg) deliquated with normal saline.Control group rats were treated with the same way of normal saline as the others.Intervention group rats were injected 1 mg of the fusion protein PEP-1/HO-1 into the left iliac vein at 30 minutes before the administration of the paraquat.After abdominal dissection with ether anaesthesia,six rats respectively from each of three groups were taken tissue samples fron kidney at l st、6 th、12 th、24 th、36 th、48 th、and 72 nd hour respectively.Tissue Superoxide Dismutase (SOD) activity and Malondialdehyde (MDA) content were measured,and renal tissue was quickly remained to observe the expression of Heme Oxygenase-1 (HO-1) by the method of imnmnohistochenistry.Blood samples were collected from the abdominal aorta fur determination of Blood Urea Nitrogen (BUN) and Creatinine (Cr) concentrations in plasma.Results To prepare high purity fusion protein PEP-1/HO-1 Successfully.The levels of the plasma Bun and Cr in Poisoned and intervention group were significantly higher than that in Control group in the six to seventy-two hours (P＜0.01);Whereas the increases of Bun andi Cr were markedly intibited in PEP-1/HO-1 intervention group,in which,the levels of Bun and Cr in the six to seventytwo hours were significantly lower than Poisoned group (P＜0.05).Comnpared with control group,the levels of MDA in poisoned groups were increased in the one to forty-eight hours and the levels of it in groups PEP-1/HO-1 in the one to thirty-six(P＜0.01 or P＜0.05).The T-SOD activity of the poisoued group was decreased in the one to thirty-six hours than that of the control group (P＜0.05).Compared with poisoned group,the T-SOD activity at various points in time was increased in group PEP-1/HO-1 (P＜0.01).In control group,there was only very.weak expressions of HO-1 in the normal renal tissue.In poisoned and intervention group,the expressions of HO-1 were significantly higher than that in control group in all time (P＜0.01).The HO-1 expressions of PEP-1/HO-1 group in all time was significantly higher than that in poisoned group (P＜0.01 or P＜0.05).Conclusion The HO-1 protein Can be successfully transduced into renal tissue by cell penetrating peptide PEP-1 and the transduced HO-1 protein reduces renal injury of the acute paraquat-induced rats by inhibiting lipid peroxidation response.

Objective:To explore the effects of bariatric metabolic surgery on body composition.Methods:The retrospective cohort study was conducted. The clinicopathological data of 66 patients with metabolic diseases who were admitted to the Third Xiangya Hospital of Central South University from January 2013 to December 2014 were collected. There were 42 males and 24 females, aged (40±11)years, with a range from 17 to 63 years. Of the 66 patients, 27 undergoing laparoscopic sleeve gastrectomy (LSG) and 39 undergoing laparoscopic Roux-en-Y gastric bypass (LRYGB) were allocated into LSG group and LRYGB group, respectively. The body composition of all patients was determined by dual-energy X-ray absorptiometry at preoperation and postoperative 6 months. Observation indicators: (1) the changes of anthropometric parameters, glucolipid metabolism, body fat mass percentage (BF%) and the ratio of Android BF% and Gynoid BF% (A/G ratio) from preoperation to postoperative 6 months; (2) the changes of whole and local body composition from preoperation to postoperative 6 months; (3) analysis of the correlation between BF% and anthropometric parameters, glucolipid metabolism. (4) Follow-up. Follow-up was conducted using outpatient or hospitalization examination to detect the changes of body composition at the time of postoperative 6 month. The follow-up time was up to July 2015. Measurement data with normal distribution were represented as Mean± SD, paired-samples t test was used for intra-group comparison, and independent-samples t test when baseline data were consistency or covariance analysis when baseline data were not consistency was used for inter-group comparison. Measurement data with skewed distribution were represented as M ( P25, P75), and comparison between groups was analyzed using Wilcoxon signed rank test. The correlation test was undertaken with the Pearson bivariate analysis. Results:(1) The changes of anthropometric parameters, glucolipid metabolism, BF% and A/G ratio from preoperation to postoperative 6 months: for patients in the LSG group, the body mass, body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), diastolic blood pressure (DBP), systolic blood pressure (SBP), fasting plasma glucose (FPG), HbA1c, high density lipoprotein cholesterol (HDL-C), triglyceride (TG), whole BF%, arms BF%, legs BF%, trunk BF%, Android BF%, Gynoid BF% and A/G ratio at preoperation and postoperative 6 months were (102±17)kg, (37±5)kg/m 2, (118±14)cm, 1.01±0.06, (94±14)mmHg(1 mmHg=0.133 kPa), (137±15)mmHg, (8.1±4.2)mmol/L, 7.3%±2.4%, (1.11±0.26)mmol/L, 2.14 mmol/L(1.73 mmol/L, 2.59 mmol/L), 40%±6%, 46%±10%, 36%±8%, 42%±6%, 45%±6%, 37%±7%, 1.23±0.18 and (82±15)kg, (29±4)kg/m 2, (101±13)cm, 0.95±0.08, (76±10)mmHg, (118±16)mmHg, (7.2±1.2)mmol/L, 5.4%±0.8%, (1.26±0.32)mmol/L, 1.21 mmol/L(0.88 mmol/L, 1.55 mmol/L), 36%±8%, 41%±9%, 34%±10%, 38%±8%, 41%±8%, 35%±10%, 1.20±0.17, respectively. There was no significant difference in the intra-group comparison of the Gynoid BF% and A/G ratio ( t=1.903, 1.730, P>0.05) and there were significant differences in the intra-group comparison of the rest of above indicators ( t=12.748, 13.283, 9.013, 3.804, 6.031, 6.226, 2.393, 4.287, -2.900, 3.193, 2.932, 5.198, 2.167, 3.357, 3.116, P<0.05). For patients in the LRYGB group, the body mass, BMI, WC, WHR, DBP, SBP, FPG, HbA1c, HDL-C, TG, whole BF%, arms BF%, legs BF%, trunk BF%, Android BF%, Gynoid BF% and A/G ratio at preoperation and postoperative 6 months were (80±12)kg, (28±4)kg/m 2, (98±9)cm, 0.96±0.05, (85±10)mmHg, (134±17)mmHg, (8.6±2.8)mmol/L, 8.3%±1.7%, (1.13±0.26)mmol/L, 2.06 mmol/L(1.15 mmol/L, 3.30 mmol/L), 30%±8%, 29%±11%, 23%±9%, 37%±7%, 40%±7%, 29%±8%, 1.42±0.26 and (69±9)kg, (24±3)kg/m 2, (91±8)cm, 0.93±0.05, (80±9)mmHg, (129±18)mmHg, (7.4±1.8)mmol/L, 7.0%±1.5%, (1.18±0.29)mmol/L, 1.29 mmol/L(0.85 mmol/L, 2.02 mmol/L), 25%±8%, 23%±12%, 20%±9%, 29%±9%, 32%±10%, 25%±9%, 1.29±0.25, respectively. There was no significant difference in the intra-group comparison of the SBP and HDL-C ( t=1.733, -1.073, P>0.05) and there were significant differences in the intra-group comparison of the rest of above indicators ( t=10.525, 10.200, 7.129, 2.887, 2.805, 2.517, 3.699, 2.608, 7.997, 8.018, 6.029, 8.342, 8.069, 5.813, 6.391, P<0.05). There were significant differences in DBP, SBP, HbA1c, trunk BF%, Android BF% and A/G ratio at postoperative 6 months between LSG group and LRYGB group ( F=6.408, t=2.641, F=20.673, 5.140, 5.735, 4.714, P<0.05). (2) The changes of whole and local body composition from preoperation to postoperative 6 months: for patients in the LSG group, the whole fat mass, muscle mass, fat-free mass at preoperation and postoperative 6 months were (38.74±9.68)kg, (57.71±11.62)kg, (60.14±11.95)kg and (26.64±8.29)kg, (48.65±13.80)kg, (51.00±14.27)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=5.256, 5.413, 5.315, P<0.05); the arms fat mass, muscle mass, fat-free mass were (5.19±1.67)kg, (5.78±1.58)kg, (6.10±1.64)kg and (3.73±1.19)kg, (5.10±1.53)kg, (5.43±1.57)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=7.564, 5.405, 5.363, P<0.05); the legs muscle mass and fat-free mass were (19.05±4.19)kg, (19.93±4.35)kg and (15.93±4.71)kg, (16.81±4.87)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=5.623, 5.568, P<0.05); the trunk fat mass and fat-free mass were (21.93±4.90)kg, (29.7±5.94)kg and (14.69±4.79)kg, (24.78±7.02)kg respectively, showing significant differences in the intra-group comparison of the above indicators ( t=8.903, 5.421, P<0.05); the Android fat mass and fat-free mass were (4.16±1.19)kg, (5.01±1.12)kg and (2.57±0.90)kg, (3.83±1.20)kg respectively, showing significant differences in the intra-group comparison of the above indicators ( t=8.288, 7.637, P<0.05); the Gynoid fat mass and fat-free mass were (5.51±1.42)kg, (9.27±1.86)kg and (3.85±1.16)kg, (7.65±2.31)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=7.461, 5.672, P<0.05); the skeletal muscle index were (8.86±1.38)kg/m 2 and (7.49±1.71)kg/m 2, respectively, showing a significant differences in the intra-group comparison ( t=5.724, P<0.05). For patients in the LRYGB group, the whole fat mass, muscle mass, bone mineral content, fat-free mass at preoperation and postoperative 6 months were (23.58±7.80)kg, (51.76±8.35)kg, (2.55±0.48)kg, (54.31±8.63)kg and (16.88±6.86)kg, (49.41±7.70)kg, (2.47±0.50)kg, (51.88±8.05)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=9.001, 3.974, 4.354, 4.075, P<0.05); the arms fat mass were (2.72±2.37)kg and (1.73±1.02)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=3.470, P<0.05); the legs fat mass, muscle mass, fat-free mass were (5.21±2.46)kg, (16.68±3.50)kg, (17.60±3.66)kg and (4.01±2.12)kg, (15.63±2.90)kg, (16.54±3.05)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=6.592, 3.372, 3.319, P<0.05); the trunk fat mass were (14.87±4.11)kg and (10.38±4.00)kg, respectively, showing a significant difference in the intra-group comparison of the above indicators ( t=8.431, P<0.05); the Android fat mass and fat-free mass were (2.61±0.86)kg, (3.96±0.87)kg and (1.81±0.79)kg, (3.78±0.67)kg respectively, showing significant differences in the intra-group comparison of the above indicators ( t=8.032, 2.153, P<0.05); the Gynoid fat mass and fat-free mass were (3.14±1.17)kg, (7.89±1.58)kg and (2.44±0.96)kg, (7.43±1.26)kg, respectively, showing significant differences in the intra-group comparison of the above indicators ( t=6.112, 3.207, P<0.05); the skeletal muscle index were (8.04±1.22)kg/m 2 and (7.43±1.13)kg/m 2, respectively, showing significant differences in the intra-group comparison ( t=4.953, P<0.05). There were significant differences in whole muscle mass, whole fat-free mass, arms fat mass, legs muscle mass, legs fat-free mass, trunk fat-free mass, Android fat-free mass, Gynoid fat-free mass and skeletal muscle index at postoperative 6 months between LSG group and LRYGB group ( F=13.846, 13.614, 23.696, 7.100, 7.127, 15.243, 16.921, 8.625, 5.497, P<0.05). (3) Analysis of the correlation between BF% and anthropometric parameters, glucolipid metabolism: the whole BF% of 66 patients was positively correlated with body mass, BMI, WC and WHR ( r=0.405, 0.663, 0.625, 0.331, P<0.05); the arms BF% was positively correlated with body mass, BMI, WC and WHR ( r=0.432, 0.682, 0.639, 0.309, P<0.05); the legs BF% was positively correlated with body mass, BMI and WC ( r=0.366, 0.646, 0.564, P<0.05); the trunk BF% was positively correlated with body mass, BMI, WC and WHR ( r=0.332, 0.560, 0.554, 0.335, P<0.05); the Android BF% was positively correlated with body mass, BMI, WC and WHR ( r=0.327, 0.537, 0.543, 0.336, P<0.05); the Gynoid BF% was positively correlated with BMI and WC ( r=0.561, 0.488, P<0.05), and negatively correlated with FPG ( r=-0.491, P<0.05); the A/G ratio was negatively correlated with BMI ( r=-0.334, P<0.05), and positively correlated with FPG ( r=0.506, P<0.05); the skeletal muscle index was positively correlated with body mass, BMI, WC and WHR ( r=0.757, 0.641, 0.609, 0.519, P<0.05), and negatively correlated with HDL-C ( r=-0.369, P<0.05). (4) Follow-up: 66 patients were followed up at the time of postoperative 6 month. Conclusions:Both LSG and LRYGB significantly change body composition. LRYGB is superior to LSG in reducing trunk BF% and Android BF%. The effects of the two surgical methods on fat mass and bone mineral content are similar. LSG lead to a more significant decrease in whole muscle mass, and LRYGB lead to a more significant decrease in legs muscle mass and skeletal muscle index.

Objective To test the effect of metastasis associated in lung adenocarcinoma transcript 1 ( MALAT1) and／or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods We transfected HCC827 cells with LV?vector or LV?over／MALAT1. Stable transfected cells ( HCC827／Vector, HCC827／MALAT1) were selected by adding puromycin. HCC827／MALAT1 cells were further transfected with the shRNA?negative control ( NC) or shRNA?human epidermal growth factor receptor 3 ( ERBB3 ) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib on the proliferation of HCC827 cells were evaluated by 3?(4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and／or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib treatment were detected by western blot. Results The MTT assay showed that sensitivity to osimertinib of HCC827／MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC50 ) of osimertinib ＞4 000 nmol／L in HCC827／MALAT1 cells.However, knockdown of ERBB3 facilitated the anti?proliferation effect of osimertinib, and the IC50 of osimertinib in shRNA?ERBB3 cells was (17.27±3.21) nmol／L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827／MALAT1 cells induced by 10 nmol／L osimertinib was (8.38±0.92)%, significantly lower than ( 27.17± 5.83)% of knockdown of ERBB3 ( P＜0.01). Western blotting showed that the expression of p?ERBB3, p?AKT and p?extracellular regulated protein kinases ( ERK) in HCC827／MALAT1 cells was markedly up?regulated, while the expression of p?epithelial growth factor receptor (EGFR) was inhibited. The expressions of p?ERBB3, p?AKT and p?ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p?EGFR, p?ERBB3, p?AKT and p?ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3／PI3K／AKT and ERBB3／MAPK／ERK signaling pathways.

Objective To investigate the relationship of procollagen?lysine 2?oxoglutarate 5?dioxygenase 2 ( PLOD2 ) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT?PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan?Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U?2OS cells with LV?vector, LV?over／PLOD2, sh?NC and sh?PLOD2. The expression of PLOD2 was detected by qRT?PCR. The impact of POLD2 on U?2OS cell invasion was determined by wound?healing assay and Transwell migration assay. The expressions of PLOD2／FAK／JAK2?STAT3 signal pathway related proteins were detected by western blotting. Results The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues ( P＜0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P＜0.01). The same results were also observed in qRT?PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 ( P＜0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+9.63)%, significantly higher than ( 9.67± 1.28)% in tissue with low expression of PLOD2 (P＜0.001).The result of wound?healing and Transwell migration assay showed that over?expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U?2OS cells ( both P＜0.01).The result of western blotting showed that over?expression of PLOD2 significantly increased the expression levels of p?FAK, p?JAK2, p?STAT3, but knockdown PLOD2 decreased the levels of p?FAK, p?JAK2, p?STAT3 in U?2OS cells. Conclusions Up?regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK／JAK2?STAT3 signal pathway.

Objective To test the effect of metastasis associated in lung adenocarcinoma transcript 1 ( MALAT1) and／or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods We transfected HCC827 cells with LV?vector or LV?over／MALAT1. Stable transfected cells ( HCC827／Vector, HCC827／MALAT1) were selected by adding puromycin. HCC827／MALAT1 cells were further transfected with the shRNA?negative control ( NC) or shRNA?human epidermal growth factor receptor 3 ( ERBB3 ) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib on the proliferation of HCC827 cells were evaluated by 3?(4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and／or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib treatment were detected by western blot. Results The MTT assay showed that sensitivity to osimertinib of HCC827／MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC50 ) of osimertinib ＞4 000 nmol／L in HCC827／MALAT1 cells.However, knockdown of ERBB3 facilitated the anti?proliferation effect of osimertinib, and the IC50 of osimertinib in shRNA?ERBB3 cells was (17.27±3.21) nmol／L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827／MALAT1 cells induced by 10 nmol／L osimertinib was (8.38±0.92)%, significantly lower than ( 27.17± 5.83)% of knockdown of ERBB3 ( P＜0.01). Western blotting showed that the expression of p?ERBB3, p?AKT and p?extracellular regulated protein kinases ( ERK) in HCC827／MALAT1 cells was markedly up?regulated, while the expression of p?epithelial growth factor receptor (EGFR) was inhibited. The expressions of p?ERBB3, p?AKT and p?ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p?EGFR, p?ERBB3, p?AKT and p?ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3／PI3K／AKT and ERBB3／MAPK／ERK signaling pathways.

Objective To investigate the relationship of procollagen?lysine 2?oxoglutarate 5?dioxygenase 2 ( PLOD2 ) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT?PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan?Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U?2OS cells with LV?vector, LV?over／PLOD2, sh?NC and sh?PLOD2. The expression of PLOD2 was detected by qRT?PCR. The impact of POLD2 on U?2OS cell invasion was determined by wound?healing assay and Transwell migration assay. The expressions of PLOD2／FAK／JAK2?STAT3 signal pathway related proteins were detected by western blotting. Results The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues ( P＜0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P＜0.01). The same results were also observed in qRT?PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 ( P＜0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+9.63)%, significantly higher than ( 9.67± 1.28)% in tissue with low expression of PLOD2 (P＜0.001).The result of wound?healing and Transwell migration assay showed that over?expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U?2OS cells ( both P＜0.01).The result of western blotting showed that over?expression of PLOD2 significantly increased the expression levels of p?FAK, p?JAK2, p?STAT3, but knockdown PLOD2 decreased the levels of p?FAK, p?JAK2, p?STAT3 in U?2OS cells. Conclusions Up?regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK／JAK2?STAT3 signal pathway.

Objective: To investigate the effect and mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1, (LncRNA-MALAT1) on invasion and metastasis of esophageal cancer cell EC-109. Methods: EC-109 cells were transfected with lentiviral vector carrying short hairpin RNA of MALAT1( shRNA-MALAT1) or a nonspecific shRNA control (shRNA-control). The expressions of MALAT1, microRNA-200a, ZEB1 and ZEB2 were detected by qRT-PCR. The effect of shRNA-MALAT1 on invasion of EC-109 cells was determined by transwell assay. The expressions of components of epithelial-msenchymal transition pathway in EC-109 cells were determined by immunofluorescence array and western blotting. The expression relationship between MALAT1 and miR-200a in EC-109 cells was detected by dual-luciferase reporter assay. Results: The result of qRT-PCR showed that the expressions levels of MALAT1, ZEB1 and ZEB2 in shRNA-MALAT1 group were 0.43±0.06, 0.64±0.04 and 0.51±0.04, respectively, significantly lower than 0.97±0.08, 1.06±0.07 and 0.98±0.05 in shRNA-control group and 1 in control group, respectively(all P<0.05). Transwell assay showed that the number of invaded cells in shRNA MALAT1 group was (96.81±10.43) per low-power field, markedly lower than that of (278.44±13.28) per low-power field in shRNA-control group (P<0.01). Immunofluorescence staining and Western blotting showed that MALAT1 downregulation significantly reduced the expressions of proteins related to EMT signal pathway in EC-109 cells.Dual luciferase reporter assay showed that compared to negative control, the activities of luciferase reporter in EC-109 cells co-transfected with pmirGLO-MALAT1-wt and miR-200a were significantly down-regulated. While co-transfected pmirGLO-MALAT1-mut with miR-200a mimics had no effect on the luciferase reporter activities of MALAT1. Conclusion LncRNA MALAT1 functions as a competing endogenous RNA to regulate the expressions of ZEB1 and ZEB2 by sponging miR-200a and promotes invasion and migration of esophageal cancer cells through inducing epithelial-mesenchymal transition.