Objective To investigate the role of Stathmin in pancreatic cancer invasion and metastasis and its relationship with DNA methylation. Methods Immunohistochemical detection of MBDI and Stathmin protein expression in 40 cases of pancreatic cancer and 15 cases ot normal pancreatic tissue were performed,followed by analysis of their clinical and pathological relationship with pancreatic cancer; Human pancreatic cancer cell line BxPC-3 was treated with 5-Aza-2-dC (AZA).Both qRT-PCR and Western blot analysis of Stathmin expression were used before and after AZA treatment; Stathmin-siRNA transfected BxPC-3 cells were divided into the Stathmi-siRNA group and the empty vector control group.Transwell chamber invasion assay and animal experiment were performed to measure the changes in cell invasion and metastatic capability. Results lmmunohistochemistry showed positive MBDI and Stathmin expressions in 28 (70％) and 24 (60％) out of 40 cases of pancreatic cancer,respectively,which were significantly higher than that in the normal pancreatic tissue (P＜ 0.05); MBDI and Stathmin protein expressions were positively correlated (r =0.356,P =0.037),so were MBDI expression and lymph node metastasis (P=0.023).Stathmin expression was significantly correlated with clinical staging and lymph node metastasis (P =0.002,and P =0.001,respectively).After AZA treatment,both Stathmin mRNA and protein expression in BxPC-3 were significantly decreased.Transwell chamber invasion assay showed that compared with the control group,the cell invasion capability of the Stathmin-siRNA group was significantly decreased (P＜0.05).Animal experiment showed that the incidence of liver metastasis was significantly lower in the Stathmin-siRNA transfected group than the empty vector control group (P＜0.05).Conclusion Demethylation may contribute to the reduction of Stathmin expression in pancreatic cancer and further improve the prognosis of pancreatic cancer patients.

Objective To evaluate the effects of celecoxib on tumor growth,COX-2 and survivin expressions and angiogenesis in nude mice. Methods Xenograft animal model was established by injecting human colon cancer HT-29 cells into the BALB/c nude mice subcutaneously. Fifty mice were randomly divided into 4 groups 16 d after injection:control group,celecoxib group(receiving 25,50,75,100 mg?kg-1?d-1 for 35 d). Tumor volumes were measured every week. The expression level of COX-2,survivin and the microvessel density (MVD) of the xenograft tumor tissues were measured by immunohistochemistry,and mRNA level of VEGF by RT-PCR. Results Celecoxib at dose of 25,50,75 and 100 mg?kg-1?d-1 inhibited the tumor volume by 34.94%,39.20%,53.50%,59.20% respectively,and showed more effective in suppressing the tumor growth than the control group(P

AIM:To establish a screening model based on muscarinic receptor type 1(M1) for drugs against Alzheimer's disease (AD). METHOD: Human M1 receptor was expressed in HEK293 (human embryonic kidney) cells, and its activation was measured by the intracellular cAMP (cyclic AMP) level. Exogeneous 3H-cAMP was used to compete the intracellular cAMP binding sites. Acetylcholine chloride was used as a positive drug to ensure the sensitivity of this model. RESULT: HEK293 cell expressing system of human M1 receptor was established. Different concentrations of acetylcholine chloride (10-9～10-4 mol/L) activation of M1 receptor leads to an increase of intracellular cAMP 10.343×10-4～33.754×10-4 pmol/μl. CONCLUTION:This screening model has positive response to M1 receptor agonist and can be used for drug screening.

Background and purpose:B-cell translocation gene 1(BTG1) can inhibit cell proliferation, promote cell apoptosis and regulate cell cycle progression and differentiation in a variety of cell types. This study aimed to explore the inlfuence on cell proliferation, apoptosis and cell cycle and its related mechanism of laryngeal cancer Hep - 2 cell lines through BTG1 overexpression byin vitro experiments.Methods:The BTG1 expression plasmids were constructed and transfected into Hep-2. They were divided into experimental group (transfected BTG1 of Hep-2 cells) and control group (transfected empty plasmid of Hep-2 cells). Western blot method was used to identify BTG1 protein expression levels of cells; proliferation activity of cells was detected by MTT assay; lfow cytometry was used to analyze the cell cycle distribution and AnnexinⅤ-FITC/PI cell apoptosis; Western blot was also used to assay cell cycle regulatory protein and apoptosis-related protein expression.Results:The pEGFP-N1-BTG1 plasmid was constructed successfully, and the expression of BTG1 protein was higher in experimental group than that in control group (0.921±0.091vs 0.308±0.047,P<0.05). Compared with the two group of laryngeal cancer Hep-2 cells, the cell growth in experimental group was slowed down and the proliferation was reduced (P<0.05); Cyclin D1 protein expression level was decreased (0.436±0.023vs 0.916±0.092,P<0.05), the proportion of G0/G1 phase cell cycle was increased [(85.1±5.2)%vs (63.8±3.1)%,P<0.05], the proportion of S phase cell was decreased [(8.3±1.1)%vs (23.1±1.5)%, P<0.05], phosphatidylserine ectropion in experimental group was increased, cell early apoptosis was significant [(10.3±1.1)%vs (2.8±0.3)%,P<0.05] and anti-apoptotic protein Bcl-2 expression level was reduced(0.167±0.009vs 0.834±0.084,P<0.05).Conclusion:BTG1 high expression could inhibit the proliferation growth of laryngeal Hep-2 cells and promote its apoptosis, and the possible mechanisms are interrelated with BTG1 involved in cell cycle regulation and causing cell apoptosis.

OBJECTIVETo study effective method of showing the microvascular architecture of human and rabbit skin flaps.

METHODSAccording to endogenous alkaline phosphatase activity, Gomori-Takamatsu method and NBT/BCIP method were used to observe the microvascular architecture.

RESULTSBoth methods had capability to demonstrate the microvascular architecture of the human skin flap, whereas the examining results of rabbits were negative.

CONCLUSIONSGreat difference of endogeneous alkaline phosphatase activity exists in different animals. Gomori-Takamatsu method and NBT/BCIP method are useful in study of microvascular architecture of human skin flaps.

Alkaline Phosphatase ; metabolism ; Animals ; Humans ; Male ; Microcirculation ; anatomy & histology ; Rabbits ; Surgical Flaps ; blood supply

Objective:To establish a colon cancer cell line which overexpressing LINC01018 stably and study its biological characteristics.Methods:The expression of LINC01018 in HCoEpiC and HT-29 cells were detected by real time fluorescence quantitative polymerase chain reaction (qRT-PCR). HT-29 cells were infected with LINC01018 overexpression lentivirus to screen and establish HT-29 cell lines which overexpressing LINC01018 stably. The effect of LINC01018 on the proliferation, invasion and migration of HT-29 cells were detected by cell counting kit-8 (CCK-8) assay and Transwell assay separately. The expression of CDK6 and matrix metalloproteinase-2 (MMP-2) in HT-29 cells was detected by Western blot.Results:The expression of LINC01018 in HT-29 cells was significantly lower than that in the human colonic epithelial cells (HCoEpiC). HT-29-L18 cell lines which overexpressing LINC01018 stably was screened successfully. Overexpression of LINC01018 significantly inhibited the cell proliferation, invasion and migration, and reduced the protein expression of CDK6 and MMP-2 in HT-29 cells.Conclusions:The expression of LINC01018 was decreased abnormally in colon cancer cells. Up-regulation of LINC01018 expression can inhibit the proliferation, invasion and migration of colon cancer cells, which may be related to CDK6 and MMP-2.

Objective:To investigate the correlation between brain iron deposition and cognitive function in patients with carotid atherosclerosis stenosis (CAS) based on quantitative susceptibility mapping (QSM).Methods:This single-center prospective study was performed at the Department of Vascular Surgery, Nanjing Drum Tower Hospital from January 2022 to June 2022. Patients who met the ataxation criteria were divided into the CAS group ( n=16) and the CAS with mild cognitive impairment (CAS-MCI) group ( n=17) according to the Montreal Cognitive Assessment (MoCA) scores. All patients completed QSM imaging and whole-brain analyses were performed for absolute susceptibility values in cortical regions. Age, sex, education years, hypertension, and diabetes mellitus were included as covariates in all analyses. Partial correlation analyses were used to determine the correlation between bilateral CAS degrees and cortical susceptibility values. Further, mediation analyses were performed to determine whether and how cortical susceptibility values affect cognition in CAS patients. Receiver operating characteristic (ROC) curve analysis was also performed to evaluate the predictive worth of differential brain region susceptibility values for cognitive decline. Independent sample t test and Mann-Whitney U test was used to compare quantitative variables. The comparison of categorical variables was conducted using χ2 test, Fisher′s exact test or Wilcoxon rank sum test. Results:A total of 33 patients were included in the study, including 16 in the CAS group and 17 in the CAS-MCI group. There were 23 males and 10 females, aged (62.8±9.0) years (range: 48 to 88 years). CAS-MCI group showed higher right CAS grades ( Z=-2.037, P=0.042). Whole-brain cortical QSM analyses showed higher susceptibility values in the frontal pole ((-0.210±0.080)×10 -8vs.(-0.130±0.120)×10 -8; t=-2.187, P=0.037), superior frontal gyrus ((-0.604±0.243)×10 -8vs. (-0.428±0.203)×10 -8; t=-2.223, P=0.034), and temporal pole ((-0.081±0.115)×10 -8vs. (0.054±0.190)×10 -8; t=-2.417, P=0.022) in CAS-MCI group compared to CAS group. The susceptibility value of the frontal pole showed a positive correlation with the right CAS grade ( r=0.424, P=0.009),while a quasi-significant positive correlation with the left CAS ( r=0.313, P=0.070). The susceptibility values of the frontal and temporal poles were negatively correlated with the MoCA score (frontal pole: r=-0.391, P=0.027; temporal pole: r=-0.410, P=0.020). Mediation analysis showed the effect of right CAS on cognition was fully mediated by the susceptibility value of the frontal pole. The ROC curve revealed that the area under the curve of using hypertension combined with the susceptibility value of the frontal pole to predict cognitive decline was 0.882 (95% CI:0.763 to 0.989) with 82% of sensitivity and 83% of specificity. Conclusions:Multiple cortical regions show iron deposition in CAS-MCI patients. Right CAS plays an important role in cognitive decline, frontal pole iron deposition mediates the effect of right CAS on cognitive function. Quantified frontal pole susceptibility is useful for the diagnosis of cognitive decline in patients with CAS.

Objective:To investigate the correlation between brain iron deposition and cognitive function in patients with carotid atherosclerosis stenosis (CAS) based on quantitative susceptibility mapping (QSM).Methods:This single-center prospective study was performed at the Department of Vascular Surgery, Nanjing Drum Tower Hospital from January 2022 to June 2022. Patients who met the ataxation criteria were divided into the CAS group ( n=16) and the CAS with mild cognitive impairment (CAS-MCI) group ( n=17) according to the Montreal Cognitive Assessment (MoCA) scores. All patients completed QSM imaging and whole-brain analyses were performed for absolute susceptibility values in cortical regions. Age, sex, education years, hypertension, and diabetes mellitus were included as covariates in all analyses. Partial correlation analyses were used to determine the correlation between bilateral CAS degrees and cortical susceptibility values. Further, mediation analyses were performed to determine whether and how cortical susceptibility values affect cognition in CAS patients. Receiver operating characteristic (ROC) curve analysis was also performed to evaluate the predictive worth of differential brain region susceptibility values for cognitive decline. Independent sample t test and Mann-Whitney U test was used to compare quantitative variables. The comparison of categorical variables was conducted using χ2 test, Fisher′s exact test or Wilcoxon rank sum test. Results:A total of 33 patients were included in the study, including 16 in the CAS group and 17 in the CAS-MCI group. There were 23 males and 10 females, aged (62.8±9.0) years (range: 48 to 88 years). CAS-MCI group showed higher right CAS grades ( Z=-2.037, P=0.042). Whole-brain cortical QSM analyses showed higher susceptibility values in the frontal pole ((-0.210±0.080)×10 -8vs.(-0.130±0.120)×10 -8; t=-2.187, P=0.037), superior frontal gyrus ((-0.604±0.243)×10 -8vs. (-0.428±0.203)×10 -8; t=-2.223, P=0.034), and temporal pole ((-0.081±0.115)×10 -8vs. (0.054±0.190)×10 -8; t=-2.417, P=0.022) in CAS-MCI group compared to CAS group. The susceptibility value of the frontal pole showed a positive correlation with the right CAS grade ( r=0.424, P=0.009),while a quasi-significant positive correlation with the left CAS ( r=0.313, P=0.070). The susceptibility values of the frontal and temporal poles were negatively correlated with the MoCA score (frontal pole: r=-0.391, P=0.027; temporal pole: r=-0.410, P=0.020). Mediation analysis showed the effect of right CAS on cognition was fully mediated by the susceptibility value of the frontal pole. The ROC curve revealed that the area under the curve of using hypertension combined with the susceptibility value of the frontal pole to predict cognitive decline was 0.882 (95% CI:0.763 to 0.989) with 82% of sensitivity and 83% of specificity. Conclusions:Multiple cortical regions show iron deposition in CAS-MCI patients. Right CAS plays an important role in cognitive decline, frontal pole iron deposition mediates the effect of right CAS on cognitive function. Quantified frontal pole susceptibility is useful for the diagnosis of cognitive decline in patients with CAS.

OBJECTIVE: To investigate the correlation between the stenosis degree of superior mesenteric artery (SMA) and each artery within the scope of aorto-iliac artery in patients with lower extremity atherosclerotic occlusive disease (LEAOD). METHODS: Images of 70 patients who had undergone the aorta-iliac-femoral arteries CT angiography (CTA) examination and had a definite diagnosis of LEAOD due to intermittent claudication or resting pain admitted to Tianjin Hospital from January to December in 2017 were enrolled. The arteries in the aorta as well as iliac were surface-reconstructed, which were analyzed by advanced vascular analysis (AVA) combined with the original images, including SMA trunk, abdominal aorta (AA), left and right common iliac artery (LCIA, RCIA), left and right internal iliac artery (LIIA, RIIA), left and right external iliac artery (LEIA, REIA). The normal reference plane and the maximal stenosis plane were selected, and the stenosis rate of each artery in the reconstruction range was automatically calculated with software. The patient's imaging data were divided into groups with two methods: (1) according to the degree of SMA stenosis, the patients were divided into group I (stenosis degree ≤70%) and group II (stenosis degree > 70%); (2) LEAOD patients with different gender were respectively divided into three groups: middle-aged group (45-59 years old), pre-elderly group (60-74 years old) and elderly group (75-89 years old). The comparison between the stenosis degree of SMA and each artery within the scope of aorto-iliac artery was analyzed with Pearson simple correlation analysis. RESULTS: The incidence of SMA stenosis in all LEAOD patients was 100%. Correlation analysis showed that there was no correlation between the stenosis degree of SMA and AA, LCIA, RCIA, LIIA, RIIA, LEIA, or REIA in group I (n = 64) and group II (n = 6), respectively (r value was -0.021, 0.023, 0.023, -0.137, 0.182, -0.113, 0.141, respectively, in group I, and it was 0.020, -0.560, 0.010, 0.306, -0.204, -0.381, 0.393, respectively, in group II, all P > 0.05). In 52 male patients, there was no correlation between the stenosis degree of SMA and AA, LCIA, RCIA, LIIA, RIIA, LEIA, or REIA in middle-aged group (n = 16), pre-elderly group (n = 27) and elderly group (n = 9), respectively (r value was -0.032, 0.024, 0.324, 0.146, 0.312, 0.008, 0.344, respectively, in middle-aged group, it was -0.108, -0.116, -0.040, -0.249, -0.082, -0.052, 0.096, respectively, in pre-elderly group, and it was 0.182, 0.311, 0.400, 0.360, 0.688, 0.498, 0.406, respectively, in elderly group, all P > 0.05). In 18 female patients, there was also no correlation between the stenosis degree of SMA and above each artery within the scope of aorto-iliac artery in pre-elderly group (n = 11) and elderly group (n = 6), respectively (the r value was -0.170, 0.040, -0.019, 0.152, 0.508, 0.042, 0.456, respectively, in pre-elderly group, and it was -0.660, 0.008, -0.055, -0.056, -0.213, 0.344, 0.011, respectively, in elderly group, all P > 0.05). The correlation in middle-aged group was not analyzed because there was only 1 patient. CONCLUSIONS Although the atherosclerotic changes in LEAOD patients can affect SMA and aorto-iliac artery at the same time, there was no correlation between the stenosis degree of SMA and each artery within the scope of aorto-iliac artery which may due to the differences in the histological structure and hemodynamics among different arteries. SMA atherosclerotic stenosis and occlusion is a relatively independent disease process for LEAOD.
Aged ; Aged, 80 and over ; Computed Tomography Angiography ; Constriction, Pathologic ; Female ; Humans ; Iliac Artery ; Lower Extremity ; Male ; Mesenteric Artery, Superior ; Middle Aged

Objective: To investigate the correlation between the stenosis degree of superior mesenteric artery (SMA) and each artery within the scope of aorto-iliac artery in patients with lower extremity atherosclerotic occlusive disease (LEAOD). Methods: Images of 70 patients who had undergone the aorta-iliac-femoral arteries CT angiography (CTA) examination and had a definite diagnosis of LEAOD due to intermittent claudication or resting pain admitted to Tianjin Hospital from January to December in 2017 were enrolled. The arteries in the aorta as well as iliac were surface-reconstructed, which were analyzed by advanced vascular analysis (AVA) combined with the original images, including SMA trunk, abdominal aorta (AA), left and right common iliac artery (LCIA, RCIA), left and right internal iliac artery (LIIA, RIIA), left and right external iliac artery (LEIA, REIA). The normal reference plane and the maximal stenosis plane were selected, and the stenosis rate of each artery in the reconstruction range was automatically calculated with software. The patient's imaging data were divided into groups with two methods: ① according to the degree of SMA stenosis, the patients were divided into group Ⅰ (stenosis degree ≤70%) and groupⅡ (stenosis degree > 70%); ② LEAOD patients with different gender were respectively divided into three groups: middle-aged group (45-59 years old), pre-elderly group (60-74 years old) and elderly group (75-89 years old). The comparison between the stenosis degree of SMA and each artery within the scope of aorto-iliac artery was analyzed with Pearson simple correlation analysis. Results: The incidence of SMA stenosis in all LEAOD patients was 100%. Correlation analysis showed that there was no correlation between the stenosis degree of SMA and AA, LCIA, RCIA, LIIA, RIIA, LEIA, or REIA in group Ⅰ (n = 64) and group Ⅱ (n = 6), respectively (r value was -0.021, 0.023, 0.023, -0.137, 0.182, -0.113, 0.141, respectively, in group Ⅰ, and it was 0.020, -0.560, 0.010, 0.306, -0.204, -0.381, 0.393, respectively, in group Ⅱ, all P > 0.05). In 52 male patients, there was no correlation between the stenosis degree of SMA and AA, LCIA, RCIA, LIIA, RIIA, LEIA, or REIA in middle-aged group (n = 16), pre-elderly group (n = 27) and elderly group (n = 9), respectively (r value was -0.032, 0.024, 0.324, 0.146, 0.312, 0.008, 0.344, respectively, in middle-aged group, it was -0.108, -0.116, -0.040, -0.249, -0.082, -0.052, 0.096, respectively, in pre-elderly group, and it was 0.182, 0.311, 0.400, 0.360, 0.688, 0.498, 0.406, respectively, in elderly group, all P > 0.05). In 18 female patients, there was also no correlation between the stenosis degree of SMA and above each artery within the scope of aorto-iliac artery in pre-elderly group (n = 11) and elderly group (n = 6), respectively (the r value was -0.170, 0.040, -0.019, 0.152, 0.508, 0.042, 0.456, respectively, in pre-elderly group, and it was -0.660, 0.008, -0.055, -0.056, -0.213, 0.344, 0.011, respectively, in elderly group, all P > 0.05). The correlation in middle-aged group was not analyzed because there was only 1 patient. Conclusions Although the atherosclerotic changes in LEAOD patients can affect SMA and aorto-iliac artery at the same time, there was no correlation between the stenosis degree of SMA and each artery within the scope of aorto-iliac artery which may due to the differences in the histological structure and hemodynamics among different arteries. SMA atherosclerotic stenosis and occlusion is a relatively independent disease process for LEAOD.