Objective:To investigate the expression of nucleoporin 43 (NUP43) in hepato-cellular carcinoma tissues and its impact on prognosis of patients and proliferation and migration of hepatocellular carcinoma cells.Methods:The retrospective cohort study and experi-mental study were conducted. The clinicopathological data of 102 hepatocellular carcinoma patients who were admitted to The First Affiliated Hospital of Army Medical University from January 2008 to December 2012 were collected. There were 83 males and 19 females, aged 56(range, 19-87)years. The expression of NUP43 in hepatocellular carcinoma tissues was analyzed by immunohistochemical staining. HepG2 and SK-HEP-1 hepatocellular carcinoma cells were cultured in vitro. The Western blot was used to verify the effects of Flag-NUP43 overexpression plasmid in transfected cells. The CCK-8 and cell migration experiments were used to analyze the effect of NUP43 overexpression on HepG2 and SK-HEP-1 hepa-tocellular carcinoma cells. Measurement data of normal distribution were expressed as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data of skewed distribution were represented as M(range). Count data were expressed as absolute numbers, and comparisons between groups was conducted using the paired chi-square test. The Kaplan-Meier method was used to calculate survival time, and Log-rank test was used for survival analysis. The R 4.2.1 software was used to draw survival curves. The COX proportional hazards regre-ssion model was used for univariate and multivariate analyses. Results:(1) Expression of NUP43 in hepatocellular carcinoma and adjacent tissues, and analysis of clinicopathological characteristics of patients with high and low expression of NUP43. Results of immunohistochemical staining showed that NUP43 was mainly expressed in the cytoplasm and nuclear membrane of cells. Of 102 hepatocellular carcinoma tissue samples, there were 49 samples with low expression of NUP43 and 53 samples with high expression of NUP43. Of 102 hepatocellular carcinoma adjacent tissue samples, there were 80 samples with low expression of NUP43 and 22 samples with high expression of NUP43. There was a significant difference in the expression of NUP43 between hepatocellular carcinoma and adjacent tissues ( χ2=16.505, P<0.05). Of 102 hepatocellular carcinoma tissue samples, there were significant differences in tumor diameter, pathological grading, and intrahepatic metastasis between the patients with low expression of NUP43 and the patients with high expression of NUP43 ( χ2=5.104, 23.217, 4.169, P<0.05). (2) Survival of hepatocellular carcinoma patients and prognostic factors analysis. The follow-up time of 102 hepatocellular carcinoma patients was 17.9(range, 0.1-107.9)months. The postoperative 1-, 3-, and 5-year overall survival rates were 79.59%, 53.06% and 34.69% for the patients with low expression of NUP43, versus 52.83%, 18.87%, and 9.43% for the patients with high expre-ssion of NUP43, showing a significant difference between them ( χ2=27.071, P<0.05). Results of multi-variate analysis showed that gender, NUP43 expression, TNM staging, and pathological grading were independent influencing factors for postoperative survival in patients with hepatocellular carcinoma ( hazard ratio=1.846, 2.206, 2.040, 2.177, 95% confidence interval as 1.231-2.768, 1.419-3.429, 1.322-3.148, 1.377-3.254, P<0.05). (3) Effects of NUP43 overexpression on the proliferation and migration of HepG2 and SK-HEP-1 hepatocellular carcinoma cells. Western blot analysis showed that transfection of Flag-NUP43 overexpression plasmid significantly increased the expression of NUP43 in HepG2 and SK-HEP-1 cells. Results of CCK-8 experiment showed that after transfection with the control plasmid and Flag-NUP43 overexpression plasmid, the cell proliferation indices of HepG2 were 0.79±0.07 and 1.47±0.05, respectively, showing a significant difference between them ( t=19.402, P<0.05). After transfection with the control plasmid and Flag-NUP43 overexpression plasmid, the cell proliferation indices of SK-HEP-1 cells were 0.59±0.05 and 0.95±0.05, respectively, showing a significant difference between them ( t=15.753, P<0.05). Results of cell migration experiments showed that after transfection with the control plasmid and Flag-NUP43 overexpression plasmid, the number of cell migrations in HepG2 was 188±8 and 595±13, respectively, showing a significant difference between them ( t=46.192, P<0.05). After transfection with the control plasmid and Flag-NUP43 overexpre-ssion plasmid, the number of cell migrations in SK-HEP-1 cells were 136±10 and 447±20, respectively, showing a significant difference between them ( t=24.721, P<0.05). Conclusions:The expression of NUP43 in hepatocellular carcinoma tissues is significantly higher than that in adjacent tissues. Gender, NUP43 expression, TNM staging, and pathological grading are independent influen-cing factors for postoperative survival of hepatocellular carcinoma patients. Overexpression of NUP43 can significantly promote the proliferation and migration of HepG2 and SK-HEP-1 hepatocellular carcinoma cells.

Objective:To investigate the expression of nucleoporin 43 (NUP43) in hepato-cellular carcinoma tissues and its impact on prognosis of patients and proliferation and migration of hepatocellular carcinoma cells.Methods:The retrospective cohort study and experi-mental study were conducted. The clinicopathological data of 102 hepatocellular carcinoma patients who were admitted to The First Affiliated Hospital of Army Medical University from January 2008 to December 2012 were collected. There were 83 males and 19 females, aged 56(range, 19-87)years. The expression of NUP43 in hepatocellular carcinoma tissues was analyzed by immunohistochemical staining. HepG2 and SK-HEP-1 hepatocellular carcinoma cells were cultured in vitro. The Western blot was used to verify the effects of Flag-NUP43 overexpression plasmid in transfected cells. The CCK-8 and cell migration experiments were used to analyze the effect of NUP43 overexpression on HepG2 and SK-HEP-1 hepa-tocellular carcinoma cells. Measurement data of normal distribution were expressed as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data of skewed distribution were represented as M(range). Count data were expressed as absolute numbers, and comparisons between groups was conducted using the paired chi-square test. The Kaplan-Meier method was used to calculate survival time, and Log-rank test was used for survival analysis. The R 4.2.1 software was used to draw survival curves. The COX proportional hazards regre-ssion model was used for univariate and multivariate analyses. Results:(1) Expression of NUP43 in hepatocellular carcinoma and adjacent tissues, and analysis of clinicopathological characteristics of patients with high and low expression of NUP43. Results of immunohistochemical staining showed that NUP43 was mainly expressed in the cytoplasm and nuclear membrane of cells. Of 102 hepatocellular carcinoma tissue samples, there were 49 samples with low expression of NUP43 and 53 samples with high expression of NUP43. Of 102 hepatocellular carcinoma adjacent tissue samples, there were 80 samples with low expression of NUP43 and 22 samples with high expression of NUP43. There was a significant difference in the expression of NUP43 between hepatocellular carcinoma and adjacent tissues ( χ2=16.505, P<0.05). Of 102 hepatocellular carcinoma tissue samples, there were significant differences in tumor diameter, pathological grading, and intrahepatic metastasis between the patients with low expression of NUP43 and the patients with high expression of NUP43 ( χ2=5.104, 23.217, 4.169, P<0.05). (2) Survival of hepatocellular carcinoma patients and prognostic factors analysis. The follow-up time of 102 hepatocellular carcinoma patients was 17.9(range, 0.1-107.9)months. The postoperative 1-, 3-, and 5-year overall survival rates were 79.59%, 53.06% and 34.69% for the patients with low expression of NUP43, versus 52.83%, 18.87%, and 9.43% for the patients with high expre-ssion of NUP43, showing a significant difference between them ( χ2=27.071, P<0.05). Results of multi-variate analysis showed that gender, NUP43 expression, TNM staging, and pathological grading were independent influencing factors for postoperative survival in patients with hepatocellular carcinoma ( hazard ratio=1.846, 2.206, 2.040, 2.177, 95% confidence interval as 1.231-2.768, 1.419-3.429, 1.322-3.148, 1.377-3.254, P<0.05). (3) Effects of NUP43 overexpression on the proliferation and migration of HepG2 and SK-HEP-1 hepatocellular carcinoma cells. Western blot analysis showed that transfection of Flag-NUP43 overexpression plasmid significantly increased the expression of NUP43 in HepG2 and SK-HEP-1 cells. Results of CCK-8 experiment showed that after transfection with the control plasmid and Flag-NUP43 overexpression plasmid, the cell proliferation indices of HepG2 were 0.79±0.07 and 1.47±0.05, respectively, showing a significant difference between them ( t=19.402, P<0.05). After transfection with the control plasmid and Flag-NUP43 overexpression plasmid, the cell proliferation indices of SK-HEP-1 cells were 0.59±0.05 and 0.95±0.05, respectively, showing a significant difference between them ( t=15.753, P<0.05). Results of cell migration experiments showed that after transfection with the control plasmid and Flag-NUP43 overexpression plasmid, the number of cell migrations in HepG2 was 188±8 and 595±13, respectively, showing a significant difference between them ( t=46.192, P<0.05). After transfection with the control plasmid and Flag-NUP43 overexpre-ssion plasmid, the number of cell migrations in SK-HEP-1 cells were 136±10 and 447±20, respectively, showing a significant difference between them ( t=24.721, P<0.05). Conclusions:The expression of NUP43 in hepatocellular carcinoma tissues is significantly higher than that in adjacent tissues. Gender, NUP43 expression, TNM staging, and pathological grading are independent influen-cing factors for postoperative survival of hepatocellular carcinoma patients. Overexpression of NUP43 can significantly promote the proliferation and migration of HepG2 and SK-HEP-1 hepatocellular carcinoma cells.

Objective:To investigate the effect of treadmill training on spasticity and the expression of potassium chloride co-transporter 2 (KCC2) after blocking BDNF-TrkB signaling pathway in rats with incomplete spinal cord injury (SCI).Methods:Forty female Sprague-Dawley rats were randomly divided into a sham-operation group (Sham group), an SCI+ phosphate-buffered saline group (SCI/PBS group), an SCI-treadmill training+ PBS group (SCI-TT/PBS group), an SCI/TrkB-IgG group and an SCI-TT/TrkB-IgG group. All of the rats underwent 1 week of intrathecal catheterization, and then T 10 incomplete SCI was induced. In the Sham group the spinal cord was only exposed. Seven days later, BDNF-TrkB signaling was blocked in the SCI/TrkB-IgG and SCI-TT/TrkB-IgG groups using the TrkB-IgG. The remaining three groups were controls treated with PBS. The SCI-TT/PBS and SCI-TT/TrkB-IgG groups began exercising 7 days after the SCI and continued for 4 weeks. The spasticity in their hind limbs was assessed using the Asworth assessment and H reflex (H-max/M-max ratio). The expression of KCC2 in the distal spinal cord was detected using western blotting and immunohistochemistry. Results:After the SCI the average Ashworth spasticity grades of the four SCI groups increased significantly compared with the Sham group. The average Ashworth spasticity grade of the SCI-TT/PBS group was significantly lower than those of the SCI/PBS and SCI/TrkB-IgG groups in the 3rd through the 5th week, and the SCI-TT/PBS group′s average grade was significantly less than that of the SCI-TT/TrkB-IgG group after 4 weeks. Within 5 weeks the average H-max/M-max ratio of the Sham group remained unchanged, significantly lower than the other 4 groups′ averages. There was no significant difference in the H-max/M-max ratio among the 4 groups of injured rats within 2 weeks after the SCI, but after 3-5 weeks the average H-max/M-max ratio of the SCI-TT/PBS group was significantly lower than those of the SCI/PBS, SCI/TrkB-IgG and SCI-TT/TrkB-IgG groups. At the 4th and 5th week the average H-max/M-max ratio in the SCI-TT/TrkB-IgG group was significantly lower than that in the SCITrkB-IgG group. And after 5 weeks the average expression of KCC2 in the anterior horn of the injured spinal cord was significantly lower in the 4 SCI groups than in the Sham group. Exercise significantly increased the expression of KCC2 in the SCI-TT/PBS group, and its immune intensity and relative optical density were significantly higher than those in the SCI/PBS, SCI/TrkB-IgG and SCI-TT/TrkB-IgG groups. However, there was no significant difference between the SCI/TrkB-IgG group and the SCI-TT/TrkB-IgG group.Conclusions:Treadmill training can improve spasticity after incomplete SCI and the expression of KCC2 in the distal spinal cord, at least in rats.

An improved manufacturing process for rivastigmine(1)was developed by performing the condensation reaction of m-hydroxyacetophenone(4)with N-ethyl-N-methyl carbamoyl chloride, then Corey-Bakshi-Shibata(CBS)chiral reduction to(R)-3-(1-hydroxyethyl)phenyl ethyl(methyl)carbamate(2)and then mesylation with methanesulfonyl chloride and nucleophilic substitution with dimethylamine, respectively. To be successful, a crucial reductive process in the conversion of ketone(3)to chiralalcohol(2)had to be correctly understood and optimized via orthogonal experiment. The whole improved process was convenient for operation and purification, with completion of the synthesis of rivastigmine and an overall yield of 88%.

Objective To investigate the expression of vascular endothelial growth factor (VEGF), angiopoietin and their receptors (VEGFR and Tie2) in aging mice kidney and the possible roles in aging mice. Methods Mice were divided as follows: 4-month old group (n=6), 9-month old group (n=6), 12-month old group (n=6) and 20-month old group (n=6). Paraffin sections of the mice kidneys were stained by PAS. The density of glomerular microvascular was determined by renal perfusion with fluorescent dyes. The level of VEGF, VEGFR2 (Flk-1), Ang-1, Ang-2, Tie2 mRNA expression and protein abundance in kidney was determined by real-time PCR, immunochemistry, immunofluorescence and Western blot. Results Compared with other three groups, in the 20-manth old group, the glomerulosclerosis index (GSI) increased remarkbly (2.48±0.79 vs 0.53±0.19, 0.69±0.18, 1.50±0.70, P<0.05); the fluorescence intensity in glomeruli decreased (P<0.05). lmmunohistochemistry demonstrated that the TGF-131 level in the aging kidneys showed an increase trend in the glomerular tubulointerstitium, and especially in the glomeruli. Real-time PCR results revealed that compared with 4-month old group mice, the mRNA expression of VEGF, Flk-1, Ang-1, Ang-2, Tie2 of the other three groups decreased, the gene levels of VEGF, Flk-1, and Ang-2 fell about 90%, 50% and 80% (all P>0.05), and the gene levels of Ang-1 and Tie2 fell about 75% and 40% in 20-month-old group (all P<0.05). Western blot domonstrated that the protein abundace of VEGF, Flk-1, Ang-1, Ang-2, Tie2 also declined with aging, the protein level of VEGF, Flk-1, Ang-1, Ang-2 and Tie2 dropped by about 35%, 50%, 15%, 13% and 21% respectively in 20-month-old group as compared to 4-month-old group (all P<0.05). Expression of above 5 factors and glomerular fluorescence intensity were negatively correlated with Scr (P<0.05). Conclusions The mRNA expression and protein abundance of VEGF, Flk-1, Ang-1, Ang-2, Tie2 in mice kidneys decreases with aging. Angiogenesis regulatory factors may play important roles in aging progression of the mice kidney.