Objective To investigate the effects of Hsp90 inhibitor 17-DMAG on proliferation and apoptosis of human colon cancer cell line HT-29.Methods HT-29 cells were treated with 17-DMAG.The cell proliferation inhibition rate was evaluated by CCK-8 assay.Apoptosis of HT-29 cells by 17-DMAG was delineated by DAPI staining assay and Annexin V PI double labeling FCM was used to determine cell apoptotic rate.Furthermore,Westen blotting analysis was used to determine caspase-3 and cleaved caspase-3 protein expression.Results 17-DMAG time-dose-dependently inhibited the proliferation of HT-29 cells.After 0.1μmol/L,0.25μmol/L,0.5μmol/L,1.0μmol/L,2.5μmol/L and 5μmol/L 17-DMAG exposured for 24 hours,the cell proliferation inhibition rate was (14.36±0.95)％,(22.17± 1.15)％,(28.45±1.16)％,(35.04±1.58)％,(46.85 ±2.44)％,(57.19 ± 2.06)％ respectively,after exposured for 48 hours,the cell proliferation inhibition rate was increased to (20.80±1.17)％,(27.55 ±0.65)％,(33.33 ±1.23)％,(46.20±4.76)％,(55.45 ±4.47)％,(61.75 ±2.72) ％ respectively,after exposure for 72 hours,the cell proliferation inhibition rate was to (29.62 ± 2.27) ％,(39.19 ± 1.74)％,(44.29 ±2.00)％,(50.66 ±2.17)％,(58.84 ±3.18)％,(70.74 ±2.65)％.DAPI staining showed that HT-29 cells treated with 17-DMAG displayed chromatin condensation and nuclear fragmentation which are typical changes of apoptosis.Annexin V PI double labeling FCM showed that when HT-29 cells were exposed to 0,0.25,0.5,1.0 and 2.5(μmoL/L) 17-DMAG for 24 hours,the total apoptotic rate for 24 hours was (2.72 ±0.57)％,(5.38 ±0.46)％,(6.88 ±0.52)％,(10.44 ±0.32)％ and (17.87 ±4.66)％ respectively.(P ＜0.05).In addition,the expression of procaspase-3 decreased,while cleaved caspase-3 increased in the presence of 17-DMAG at different concentrations for 24 hours.Conclusion 17-DMAG can time-dose-dependently inhibit proliferation and promote apoptosis of HT-29 cells in vitro.

Objective To investigate the diagnostic value of 18F-FDG PET/CT features of spinal tuberculosis.Methods 18 F-FDG PET/CT imaging manifestation of 125 cases (70 males,55 females,average age 44.3 years) with osseous or mixed vertebra bone destruction from January 2006 to June 2014 were analyzed retrospectively.The patients included 32 cases with spinal tuberculosis (50 lesions,study group) and 93 cases with other diseases (150 lesions,control group).The corresponding FDG PET/CT features were analyzed by two classification logistic regression (Forward:LR),then FDG PET/CT features with statistical significance in diagnosis of spinal tuberculosis were obtained.The sensitivity and specificity of those FDG PET/CT features were calculated.Results Odds ratio (OR) of FDG PET/CT features with statistical significance in diagnosis of spinal tuberculosis from high to low were paraspinal cold abscess (20.790),cold zone(10.528),intervertebral disc involvement(5.394),continuous vertebral involvement(3.493).The diagnostic sensitivity and specificity were 22.0％ (11/50),99.3％ (149/150) forcold abscess,70.0％ (35/50),90.0％ (135/150) for cold zone,82.0％ (41/50),83.0％ (125/150) for intervertebral disc involvement,82.0％ (41/50),78.0％ (117/150) for continuous vertebral involvement,respectively.Spinal tuberculosis often had two or more signs.The continuous vertebral involvement + intervertebral disc involvement was most common,with the diagnostic sensitivity and specificity of 78.0％ (39/50) and 88.7％ (133/150).The diagnostic sensitivity and specificity of continuous vertebral involvement+intervertebral disc involvement+cold zone were 58.0％ (29/50),96.0％ (144/150),and those of four features conourrence were 14.0％ (7/50),100％ (150/150),respectively.Conclusion Regression analysis shows that FDG PET/CT features with statistical significance in diagnosis of spinal tuberculosis are paraspinal cold abscess,cold zone,intervertebral disc involvement,continuous vertebral involvement,which have high diagnostic sensitivity and specificity.

Objective To investigate the relationship between serum levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) and Suzuki' s grading system in adult moyamoya disease (MMD).Methods Fifty-two adult patients with MMD,who were diagnosed in Jinling hospital between April 2009 and January 2010,were retrieved from the Nanjing Stroke Registry Program (NSRP).Sixteen sex- and age-matched healthy individuals with MMD patients consisted of the control group.Using enzyme-linked immunosorbent assay,serum concentrations of VEGF and MMP-9 were compared between adult MMD patients and healthy individuals.By Suzuki' s six-grading system,patients were divided into different subgroups,and the correlation of serum levels of VEGF and MMP-9 corresponding to different subgroup and Suzuki's grading was respectively analyzed.In addition,the correlation of serum levels of VEGF and MMP-9 was also evaluated.Results Serum VEGF concentrations in ischemic and hemorrhagic MMD patients was respectively ( 289.4 + 69.2 ) pg/ml and ( 324.3 ± 95.6 ) pg/ml and were significantly higher compared to those in healthy controls ( ( 63.5 ± 7.6 ) pg/ml; F =69.43,P ＜ 0.01 ).Similar findings were observed for MMP-9 ( ( 499.4 ± 76.2 ) ng/ml and ( 531.2 + 100.2 ) ng/ml versus (257.1 ±30.7) ng/ml; F =66.023,P ＜0.01 ).With the increase of Suzuki' s grading,serum levels of VEGF and MMP-9 respectively showed a high trend ( r =0.879,P ＜ 0.01:r =0.838,P ＜ 0.01 ).In addition,a positive correlation between serum levels of VEGF and MMP-9 was found in the MMD group( r =0.590,P ＜0.01 ).Conclusion The results show that serum levels of VEGF and MMP-9 in adult MMD are higher than those in healthy controls,which may play a role in neovascularization in MMD,and moreover,serum levels of VEGF and MMP-9 show a high trend with the progression of MMD,which suggest that serum levels of VEGF and MMP-9 can reflect the severity of MMD.

Objective To investigate the value of hyperintense vessel signs (HVS) on fluidattenuated inversion recovery (FLAIR) sequence for assessing the patterns of collateral blood flow in adult moyamoya disease (MMD).Methods Forty-one adult patients with non-hemorrhagic MMD retrieved from Nanjing Stroke Registry Program between August 2008 and January 2011 were identified by digital cerebral angiography and performed the examination of FLAIR sequence in Jinling hospital.According to the different sites of HVS located in the territory of the middle cerebral artery,the patterns of HVS were classified into grades 0-3: Grade 0,absence of HVS ; Grade 1,HVS limited in the cerebral sulci of temporal lobe and Sylvian fissure ; Grade 2,HVS in the cerebral sulci of frontal and parietal lobe regions and Sylvian fissure;and Grade 3,HVS in the combined territories of Grade 1 and Grade 2.According to the intracerebral collateral blood flow,steno-occlusions of the arteries were classified into three types: Type 1,residual antegrade flow across steno-occlusive lesions; Type 2,retrograde flow via leptomeningeal vessels; Type 3,the combined collateral blood flow of Type 1 and Type 2.The relationship between the patterns of intracerebral collateral blood flow and the location of HVS was analyzed.Results Of 41 adult patients with non-hemorrhagic MMD,there were 3 patients presented with unilateral vascular lesions and 38 with bilateral vascular lesions,so the total number of vascular lesions of the cerebral hemispheres was 79.Because three patients showed the absence of HVS in bilateral hemispheres,the total number of the presence of HVS of the cerebral hemispheres was 73.Therefore,the percentage of the presence of HVS was 92.4％ (73/79) in vascular lesions of the cerebral hemispheres.Importantly,the patterns of slow collateral blood flow corresponding to Grade 1 HVS were all antegrade (7/7) ; the collateral patterns corresponding to Grade 2 HVS were mainly retrograde leptomeningeal flow (95.0％,19/20) ; and the patterns corresponding to Grade 3 HVS were mainly slow combined collateral blood flow(84.8％,39/46).Furthermore,with the changing sites of HVS from the cerebral sulci of temporal lobe to the cerebral sulci of frontal and parietal lobe regions,the directions of collateral flow changed with a shift from antegrade to retrograde,which was statistically significant.Conclusion The different locations of HVS can reflect the different patterns of collateral blood flow,and the locations of HVS may predict the directions of intracerebral collateral blood flow in adult MMD patients.

To develop a low-cost, reliable, easy-to-maintain and practical instrument for protein purification. Some ultraviolet luminescent diode was used to provide 280 nm light source, and high-sensitivity S1336 photo-electric detector was employed for real-time monitoring of purified protein solution flowing through quartz cell to supervise the concentration of the protein. The instrument gave voice alarm and stopped working in case the protein concentration was less than the standard one. The lower SCM monitored the liquid level of the protein collecting cup and the position of loading arm through laser infrared distance sensor, so that a cup full of protein might be replaced by another empty cup. The instrument involved in Samsung S5PV210 embedded master computer, Wince6.0 operating system, Keil4.0 and VS2005. Trials proved that the instrument could perform real-time monitoring and curve display of dual-channel ultraviolet absorption, and could realize auto collection of 735 ml protein solution and up to 5-hour standby. The instrument developed has simple structure, high reliability and easy maintenance, and meets the desired require-ments.

Objective To investigate the clinical value of susceptibility weighted imaging (SWI)combined with three dimensional pseudo continuous arterial spin labeling (3D-PCASL)on the prognosis analysis of acute cerebral infarction.Methods Thirty cases with acute cerebral infarction (< 72 h)underwent conventional MRI,MRA,3D-PCASL and SWI.NIHSS scores were performed at the time of examination and 3 months later.The correlation between the collateral blood vessels,regional cerebral blood flow (rCBF)detected by combination of SWI and 3D-PCASL with clinical prognosis were analyzed.Results Twenty-three cases showed collateral blood vessels in the lesions with 1 grade in 14,and 2 grade in 9.The average rCBFs in grade 0,1,2 infarction areas were (22.69±11.94)mL·100 g-1 ·min-1 ,(25.10±16.55)mL·100 g-1 ·min-1 and (33.04±24.24)mL·100 g-1 ·min-1 ,respectively.Collateral blood vessels,rCBF were positive correlated with the NIHSS scores (r=0.989,P< 0.01).18 cases showed multiple vessels around the lesions. The average rCBFs in the infarction area with or not with periphery collateral blood vessles were (28.33±24.24)mL·100 g-1 ·min-1 and (22.69±11.94)mL·100 g-1 ·min-1 ,respectively.There was a positive correlation between rCBF and NIHSS scores (r=0.897,P<0.01). Of 30 cases of acute cerebral infarction,the average CBFs in the infarct areas and the contralateral mirror areas were (26.92±18.22)mL·100 g-1 · min-1 and (34.22±12.37)mL·100 g-1 ·min-1 .There was significant difference (t=8.093,P<0.01).Conclusion The combination of SWI and 3D-PCASL can display the collateral blood vessels in the lesions and soft meninges,and provide the quantitative analysis of rCBF,which has important clinical significance for prediction of the prognosis of acute cerebral infarction.

Objective To explore the reasons for rebleeding and its coping strategies.Methods To study the 19 hypertensive intracerebral hemorrhage patients'medical records who suffered postoperative rebleeding in our department from June 2011 to May 2015.Results Rebleeding time:within 6 hours:11 cases,6 ~24 hours:5 cases, 2 ~3 weeks:3 cases.Among 19 cases,hematoma puncture drainage in 2 cases,small bone window craniotomy surgery in 9 cases,craniotomy with big bone flap decompression in 8 cases,6 cases with severely disabled and 7 cases died. Conclusion Intraoperative bleeding operation is not standardized,early postoperative blood pressure control failure in patients,preoperative instability function are the main causes of postoperative bleeding.Choosing a better operating time,the specification intraoperative hemostasis and controlling blood pressure to stable after surgery can reduce the membership of hypertensive intracerebral hemorrhage incidence of postoperative bleeding.

Objective:To investigate the influencing factors of goiter in school-age children aged 8 to 10 in non-high iodine areas of Shijiazhuang City.Methods:In April 2018, 9 non-high iodine counties (cities) were selected as monitoring sites in Shijiazhuang City, and capacity proportional probability sampling (PPS) method was used. Each monitoring site was divided into five sampling areas according to five orientations: east, west, south, north, and middle, one township was selected from each area, one elementary school was selected from each township, and 40 school-age children aged 8 to 10 (balanced age, half males and half females) were selected from each school as respondents. Urine samples from any one time of children and drinking water samples from their village were collected, and urinary iodine and water iodine were detected by arsenic cerium catalytic spectrophotometry; the thyroid volume of children was measured by B ultrasound method; at the same time, the height and weight of children were measured and the body mass index was calculated. The influencing factors of goiter were analyzed by logistic regression analysis.Results:A total of 1 867 urine samples of school-age children were collected, and the median urinary iodine was 190.65 μg/L, which was in the suitable level of iodine. A total of 1 046 drinking water samples were collected, water iodine ranged from 0.11 to 87.91 μg/L, and the median water iodine was 3.01 μg/L. A total of 1 867 school-age children were tested thyroid, the median thyroid volume was 3.01 ml. The medians thyroid volume of boys and girls (928 and 939 cases) were 2.90 and 3.13 ml, respectively, the difference was statistically significant between sex ( U = 2.09, P < 0.05); the medians thyroid volume of children aged 8, 9, and 10 years old (622, 629, 616 cases) were 2.47, 2.87, and 3.13 ml, respectively, the differences were statistically significant among ages ( H = 203.96, P < 0.01); the medians thyroid volume of normal, overweight and obese children (1 231, 300, 336 cases) were 2.61, 3.05 and 3.16 ml, respectively, the differences were statistically significant among body mass index ( H = 65.55, P < 0.01). The results of multifactorial logistic regression analysis showed that female and obesity were risk factors of goiter in school-age children [odds ratio ( OR) = 2.08, 2.86, 95% confidence interval ( CI): 1.05 - 4.12, 1.39 - 5.88, P < 0.05]. Conclusion:Female and obesity are risk factors of goiter in school-age children aged 8 to 10 in non-high iodine areas of Shijiazhuang City.

Neddylation is a post-translational protein modification process. MLN4924 is a newly discovered pharmaceutical neddylation inhibitor that suppresses cancer growth with several cancer types. In our study, we first investigated the effect of MLN4924 on colon cancer cells (HCT116 and HT29). MLN4924 significantly inhibited the neddylation of cullin-1 and colon cancer cell growth in a time and dose-dependent manner. MLN4924 induced G2/M cell cycle arrest and apoptosis in HCT116 and HT29 cells. Moreover, MLN4924 also triggered autophagy in HCT116 and HT29 cells via suppressing the PI3K/AKT/mTOR pathway. Inhibiting autophagy by autophagy inhibitor 3-MA or ATG5 knockdown reversed the function of MLN4924 in suppressing colon cancer cell growth and cell death. Interestingly, MLN4924 suppresses colon cell growth in a xenograft model. Together, our finding revealed that blocking neddylation is an attractive colon cancer therapy strategy, and autophagy might act as a novel anti-cancer mechanism for the treatment of colon cancer by MLN4924.
Apoptosis ; Autophagy* ; Cell Cycle Checkpoints ; Cell Death ; Colon* ; Colonic Neoplasms* ; Heterografts ; HT29 Cells ; Humans ; Protein Processing, Post-Translational

Objective:To explore the reasonable timing of radiotherapy for epidermal growth factor receptor ( EGFR) mutation-positive non-small cell lung cancer patients with brain metastasis in the era of third-generation targeted drugs. Methods:Clinical data of EGFR mutation-positive non-small cell lung cancer patients with brain metastasis who received first-line treatment with third-generation targeted drugs and stereotactic radiotherapy at Shanghai Armed Police Corps Hospital from September 2019 to May 2022 were retrospectively analyzed. According to the timing of radiotherapy before / after targeted drug resistance, all patients were divided into the early and salvage radiotherapy groups. The proportion of brain metastasis, physical fitness, complete response rate, objective response rate, delaying the progression of brain metastasis and overall survival (OS) were compared between two groups. Kaplan-Meier method was used for survival analysis, log-rank test was used for univariate prognostic analysis, and factors with P <0.1 were included in Cox multivariate analysis. Results:A total of 85 patients were included, including 51 (60%) cases receiving early radiotherapy. Patients who participated in early radiotherapy had a higher proportion of symptomatic brain metastasis (82% vs. 56%, P=0.013) and poorer physical fitness (Kanofsky performance score <70: 61% vs. 26%, P=0.002) compared to patients who underwent salvage radiotherapy. Early radiotherapy significantly improved the complete response rate of intracranial lesions (35% vs. 12%, P=0.015) and objective response rate (88% vs. 71%, P=0.041), delayed the progression of brain metastasis (median intracranial progression free survival: 23.0 months vs. 16.0 months, P=0.005; median intracranial secondary progression free survival: 31.0 months vs. 22.0 months, P=0.021), and improved OS (median OS: 44.0 months vs. 35.0 months, P=0.046). In multivariate analysis, diagnosis-specific graded prognostic assessment score <2.5, mutation of EGFR exon 21, and salvage brain radiotherapy were adverse prognostic factors for OS. Conclusion:In the era of third-generation targeted drugs therapy, early involvement of stereotactic radiotherapy in non-small cell lung cancer patients with brain metastasis can bring greater clinical benefits.