温州医科大学学报
 
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2022 Vol. 52, No. 8
Published: 2022-08-25
 
 
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2022 Vol. 52 (8): 0- [Abstract] ( 272 ) HTML (1 KB)  PDF (4894 KB)  ( 591 )
603 Target network recognition and compatibility mechanism of Astragalus-Zedoary inducing apoptosis of cervical cancer cells
SUN Hao, ZHENG Yongquan, XU Mengjin
DOI: 10.3969/j.issn.2095-9400.2022.08.001
Objective: To identify the target network of Astragalus-Zedoary inducing apoptosis of cervical cancer cells and explore their compatibility mechanism. Methods: First, the cervical cancer disease network was constructed by bioinformatics and the background network was obtained by GO clustering, and the potential key targets were identified by K-means analysis. Then, apoptosis-related target network was identified by k-core decomposition of apoptosis-related sub-networks in the background network. Subsequently, potential key targets in the target network were evaluated by molecular docking and TOPSIS, and potential active ingredients were identified at the same time. Finally, the cell and molecular experiments such as CCK-8 method, TUNEL staining,flow cytometry and Western blot analysis were used to evaluate the ability of Astragalus-Zedoary to induce apoptosis of cervical cancer SiHa cells and to explore the potential compatibility mechanism. Results: The network pharmacology analysis showed that potential active ingredients of 64 Astragalus and 44 Zedoary synergistically regulated the target network composed of 14 potential key targets, while the vast majority of Astragalus (60) and Zedoary (38) ingredients acted on the p53 and β-catenin, respectively. The experimental results showed that the IC50 of Astragalus-Zedoary aqueous extract at 24 h and 48 h were 62.47 mg/mL and 50.37 mg/mL, respectively.Meanwhile, Astragalus-Zedoary aqueous extract could induce G0/G1 phase arrest and early apoptosis in cervical cancer SiHa cells. In addition, Astragalus-Zedoary aqueous extract could increase the expression level of p53 (P<0.05), but the effect on β-catenin is not obvious (P>0.05). Conclusion: Astragalus-Zedoary might synergistically regulate the target network dominated by p53 through its multiple active ingredients, so as to arrest the cell cycle in G0/G1 phase, thereby inducing early apoptosis of cervical cancer cells.
2022 Vol. 52 (8): 603-612 [Abstract] ( 354 ) HTML (1 KB)  PDF (2623 KB)  ( 649 )
613 Preparation, characterization of micheliolide DSPE-PEG nanomicelles and its induction of apoptosis in glioma cells
WANG Chengde, XU Yunqiu, SU Zhipeng
DOI: 10.3969/j.issn.2095-9400.2022.08.002
Objective: To prepare and characterize micheliolide-loaded polyethylene glycol-derivatized phosphatidyle-thanolamine (MCL@DSPE-PEG) nanomicelles, and evaluate its anti-tumor effect on tumor cells. Methods: MCL@DSPE-PEG nanomicelles were prepared by film hydration method, and their appearance characteristics were observed. The particle size, polydispersion coefficient (PDI), drug-loading amount and encapsulation efficiency of the nanomicelles were detected. The drug release of MCL@DSPE-PEG nanomicelles in pH 7.4 phosphate buffer were compared within 0-144 h. The U87 and U251 glioma cells were divided into model group and MCL@DSPE-PEG nanomicelles group. Cell viability and apoptosis were evaluated. The expressions of apoptosis related protein including Caspase3, Caspase8 and Bcl-2 were detected. Results: The morphology of MCL@DSPE-PEG nanomicelles were uniform ellipsoidal shape. The particle size was (123.8±0.5) nm and PDI was 0.236±0.005. The drug-loading amount was 8.96%±0.56%, encapsulation efficiency was 85.49%±3.66%. Accumulative release rate was 73.30% within 144 h. MCL raw material was released completely within 48 h. MCL@DSPE-PEG nanomicelles inhibited the growth and cell cloning of U87 and U251 glioma cells. Compared with the model group, the expression of Bcl-2 protein was significantly decreased, while the expression of Caspase3 and Caspase8 protein were dramatically increased (P<0.05 respectively). Conclusion:MCL@DSPE-PEG nanomicelles are prepared successfully, which can induce apoptosis of U87 and U251 glioma cells and inhibit tumor growth.
2022 Vol. 52 (8): 613-619 [Abstract] ( 575 ) HTML (1 KB)  PDF (1784 KB)  ( 772 )
620 Rhein attenuates hypoxia-induced pulmonary hypertension via CDK9/STAT3 pathway
CHEN Mayun, YAO Yizhu, ZHU Lin, CAI Chaoyang, XU Ziwei, SUN Junwei, WANG Liangxing, HUANG Xiaoying.
DOI: 10.3969/j.issn.2095-9400.2022.08.003
Objective: To investigate the effect of rhein on hypoxia-induced pulmonary hypertension (HPH) and its mechanism. Methods: Sprague-Daeley (SD) rats were used as the research objects. The SD rats separated into 3 groups: normoxia group (N), hypoxia group (H), hypoxia+rhein group (HR). The expressions of p-CDK9,CDK9, p-STAT3/STAT3, PCNA, Bcl-2, Bax, Caspase-3, CC-3 (Cleaved-Caspase-3) in lung homogenate were revealed by Western blot. The mean pulmonary artery pressure in 3 groups of rat was measured by right heart catheterization method. The degree of right cardiac hypertrophy was measured by weighing, while the degree of pulmonary vascular remodeling in each group of rats was observed by HE staining. Results: Compared with the N group, the p-CDK9, CDK9, p-STAT3/STAT3, PCNA, Bcl-2 protein expression in H group was increased (P<0.05), the Bax, Caspase-3, CC-3 protein expression in H group was reduced (P<0.05), the mPAP, right cardiac
hypertrophy index and WA/TA in H group were considerably enhanced (P<0.05). In contrast with the H group,the p-CDK9, CDK9, p-STAT3/STAT3, PCNA, Bcl-2 protein expression in HR group was reduced (P<0.05), the Bax, Caspase-3, CC-3 protein expression in HR group was considerably increased (P<0.05), the mPAP, right cardiac hypertrophy index and WA/TA in HR group were reduced (P<0.05). Conclusion: Rhein could inhibit the phosphorylation of STAT3 by inhibiting the expression and phosphorylation of CDK9, thereby alleviating pulmonary hypertension, improving pulmonary vascular remodeling and right heart hypertrophy in HPH rats.
2022 Vol. 52 (8): 620-624 [Abstract] ( 452 ) HTML (1 KB)  PDF (2188 KB)  ( 817 )
625 The application of drug-coated balloon in in situ and long lesions of large vessels in coronary arteries
YAN Xigui, PENG Huaiming, ZHENG Yong, HUANG Chunping, LIU Wenmin1, YANG Deye
DOI: 10.3969/j.issn.2095-9400.2022.08.004
Objective: To observe the efficacy of drug-coated balloon (DCB) in the treatment of large vessels in situ (reference diameter≥2.8 mm) and in situ long lesions (reference diameter≥2.8 mm, length≥20 mm) of large vessels in coronary arteries. Methods: A total of 80 patients who were diagnosed and treated in the Department of Cardiology, Affiliated Hospital of Hangzhou Normal University from April 2015 to December 2020 were selected, who were all treated with DCB for in situ primary coronary artery lesions. Computer quantitative coronary angiography analysis software was used to analyze the coronary angiography images of the included patients at three different times: before operation, immediately after operation and follow-up. The selected patients were divided into two groups according to the length of target vessel lesions, namely the lesion length <20 mm group (n=45) and the lesion length≥20 mm group (n=35). The inter-group comparison was performed to evaluate the efficacy of DCB in long in situ coronary artery lesions. Results: The average length of target vessel lesion of the included cases was (15.71±5.98) mm, and the reference diameters was (3.22±0.34) mm. During the follow-up of (15.89±9.34) months, no patients had acute myocardial infarction or severe arrhythmia, cardiogenic
death or stroke. There were no cases of revascularization of target vessel in all follow-up patients, and the late lumen loss during follow-up was (-0.01±0.26) mm. There was no significant difference in late lumen loss between the two groups of patients with different lesion lengths [(0.003±0.3) mm vs. (-0.03±0.23) mm, P=0.615].
Conclusion: DCB is safe and effective in the treatment of in situ and long lesions of large vessels in coronary arteries and it can reduce the late lumen loss of coronary target lesions.
2022 Vol. 52 (8): 625-631 [Abstract] ( 376 ) HTML (1 KB)  PDF (1356 KB)  ( 693 )
632 The statistical performance of different strategies for dealing with missing values in clinical trials
ZHAO Shuzhen, JIN Dongzhen, LI Huihui, LAI Mengyuan, HUANG Ruogu, MAO Guangyun.
DOI: 10.3969/j.issn.2095-9400.2022.08.005
Objective: To evaluate the statistical performance of various missing value processing methods in a two-arm superiority clinical trial with different missing patterns, mechanisms and proportions. Methods:We generated longitudinal simulation datasets containing monotonic and arbitrary missing patterns, missing mechanisms that occurred at random or completely at random, and missing proportions of 0%-5%, 5%-10% and 10%-15%, respectively. Estimations of efficacy between the two groups after performing treatment in each simulated dataset by different missing value handling methods, and the statistical performance of the different methods were evaluated according to the magnitude of the difference in estimated efficacy from the full data set.Results: When the proportion of missingness was <5%, the effect estimates obtained from different missingness treatments in any missing patterns were close to each other, and the effect estimates from repeated measures mixed effects model (MMRM) and the analysis of covariance with multiple imputations in monotonic missing patterns were closest to the true values. When the proportion of missingness >5%, the effect estimates from MMRM with different covariance matrix structures and the analysis of covariance with multiple imputations were still closest to the true values regardless of the missing mechanisms and missing patterns, with the former more stable than the latter. In contrast, the error in the effect estimates of the single imputation approach and the mixed-mode model (PMM) increased with the proportion of missing, especially for the monotonic missing, where the error was greatest when the proportion of missing was 10%-15%. Conclusion: MMRM yielded the most accurate effect estimates for different missing proportions (0%-5%, 5%-10% and 10%-15%), missing patterns (monotonic missing or arbitrary missing), and missing mechanisms (completely random missing or random missing),suggesting that MMRM is the preferred solution for dealing with missing longitudinal quantitative data in twoarm superiority clinical trials.
2022 Vol. 52 (8): 632-637 [Abstract] ( 417 ) HTML (1 KB)  PDF (1552 KB)  ( 1274 )
638 Prediction of rupture risk for middle cerebral artery aneurysm in patients with hypertension based on nomogram
SU Na, ZHOU Jiafeng, LIN Boli, ZHU Dongqin, YANG Yunjun, CHEN Yongchun
DOI: 10.3969/j.issn.2095-9400.2022.08.006
Objective: To investigate the risk factors of middle cerebral artery (MCA) aneurysm rupture in patients with hypertension, and to construct a nomogram prediction model. Methods: The clinical and imaging data of hospitalized patients with MCA aneurysms complicated with hypertension from January 2009 to June 2020 were collected retrospectively. All patients underwent CT angiography (CTA) and the morphological parameters of aneurysms were measured. Multivariate logistic regression was used to analyze the independent risk factors of MCA aneurysm rupture in patients with hypertension and a nomogram prediction model was constructed. The area under receiver operating characteristic curve (AUC) was used to evaluate the diagnostic efficiency of the model. Results: A total of 254 patients with MCA aneurysms complicated with hypertension were finally included in this study. Multivariate logistic regression showed that gender (OR=2.277, P=0.022), aspect ratio (OR=10.270,
P<0.001), irregular shape (OR=4.747, P<0.001) and location (OR=3.161, P=0.001) were independent risk factors for MCA aneurysm rupture in patients with hypertension. The nomogram prediction model has good diagnostic efficiency (AUC=0.866) and the prediction accuracy is 82.68%. Conclusion: Aspect ratio, irregular shape,location and gender are independent risk factors for MCA aneurysm rupture in patients with hypertension. It is feasible to predict the rupture risk of MCA aneurysm in patients with hypertension based on nomogram, which provides a visual basis for clinical personalized diagnosis and treatment.
2022 Vol. 52 (8): 638-644 [Abstract] ( 438 ) HTML (1 KB)  PDF (1561 KB)  ( 611 )
645 CT-based radiomics nomogram to differentiate non-functional pancreatic neuroendocrine tumors from solid pseudopapillary tumors
ZHOU Yongjin, GAO Ruijie, JIANG Chunyan, DENG Jingjing, XIA Shuiwei, SHEN Shaobo, WANG Zufei, JI Jiansong
DOI: 10.3969/j.issn.2095-9400.2022.08.007
Objective: To investigate the value of CT-radiomic model for preoperative differentiating nonfunctional pancreatic neuroendocrine tumors (NF-pNETs) from solid pseudopapillary tumors (SPTs). Methods:A total of 87 cases with pathologically confirmed NF-pNETs and SPTs were included and randomly divided into training set (62 cases) and validation set (25 cases) at a ratio of 7:3. A.K. software was used to extract tumor texture features from plain CT, arterial and delayed CT images. The maximum correlation and minimum redundancy (mRMR), least absolute shrinkage and selection operator (LASSO) and 10-fold cross validation were used to retain features. Logistic regression analysis and Rad-score were used to construct the radiomic model nomogram. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficiency of the radiomic model nomogram. Decision curve analysis (DCA) was used to evaluate the benefits of the radiomic nomogram. Results: Totally 396 features were extracted from plain CT scan, arterial phase and delay phase images respectively. Ultimately, 8, 6 and 7 features were retained after mRMR, LASSO and 10-fold cross validation respectively, and 14 features were retained for the combined radiomic models. Multivariate logistic regression was used to construct a combined radiomic model nomogram including gender, age, maximum tumor diameter and texture extracted from plain CT scan, arterial phase, and delay phase images. The combined radiomics model achieved better prediction performance than radiomics model based on the single CT plain scan, arterial phase and portal venous phase CT images. The area under curve of the combined radiomics model was 0.97 (95%CI=0.94-1.00) for the training set and 0.92 (95%CI=0.81-1.00) for the validation set. DCA revealed that when the risk threshold was greater than 0.45, the use of combined radiomics model to differentiate between the tumor and pancreas to predict SPTs vs. NFNETs was of greater clinical value than a treat-all-patients as SPTs scheme or a treat-none as NF-NET scheme. Conclusion: Radiomics nomogram based on combined plain CT scan, arterial phase and delayed phase images shows good performance to distinguish NF-pNETs from SPTs,
which may be used as a noninvasive imaging tool for preoperative evaluation.
2022 Vol. 52 (8): 645-651,656 [Abstract] ( 483 ) HTML (1 KB)  PDF (1704 KB)  ( 595 )
652 Incidence of deep venous thrombosis after intracerebral hemorrhage in the middle-aged and elderly and its influencing factors
CHEN Xiufang, SUN Zifen, WANG Xiaoxue, DONG Xiaochen, HU Pinglang,HONG Xianchai, CHEN Chan, WANG Hongxia
DOI: 10.3969/j.issn.2095-9400.2022.08.008
Objective: To investigate the incidence of acute deep vein thrombosis (DVT) after intracerebral hemorrhage in the middle-aged and elderly and explore its influencing factors, so as to provide reference for the prevention and nursing care. Methods: A total of 245 patients with cerebral hemorrhage hospitalized within 24 hours of onset in the First Affiliated Hospital of Wenzhou Medical University from September 2019 to May 2021 were selected. All patients underwent venous ultrasound examination of lower extremities 10-14 days after onset.The prevalence of DVT in the acute stage after cerebral hemorrhage was calculated. The influencing factors of DVT after intracerebral hemorrhage in middle-aged and elderly patients were analyzed by univariate analysis and binary logistic regression analysis. Results: Of 245 middle-aged and elder patients with intracerebral hemorrhage investigated, 79 cases had DVT, with the incidence rate of 32.24%, of which 39 cases (49.37%) had no clinical symptoms of DVT. Binary logistic analysis showed that age ≥70 years old (OR=1.73, 95%CI=1.12-2.98),NIHSS score of lower limbs score ≥3 points (OR=4.13, 95%CI=2.15-8.97), continuous bed time ≥3 days (OR=4.73, 95%CI=2.25-9.78), Wells ≥2 points (OR=3.54, 95%CI=1.62-6.67), and high D-dimer at admission (OR=3.06, 95%CI=1.01-3.49) were independent risk factors for DVT in the acute stage after cerebral hemorrhage.Early rehabilitation was a protective factor for DVT (OR=0.55, 95%CI=0.28-0.96). Conclusion: The incidence of DVT after intracerebral hemorrhage was high in the middle-aged and elderly, and nearly half of patients do not have clinical symptoms of DVT. Many independent risk factors are involved in DVT of the middle-aged and elderly patients with acute stroke. Timely monitor and intervention are suggested for patients with risk factors and early rehabilitation treatment of cerebral hemorrhage should be combined to reduce the occurrence of DVT.
2022 Vol. 52 (8): 652-656 [Abstract] ( 390 ) HTML (1 KB)  PDF (1341 KB)  ( 650 )
657 Effects of sodium valproate on the progression of non-alcoholic fatty liver disease by inhibiting prolyl endopeptidase activity
LI Mengting, JIANG Daixi, ZHANG Jianbin, WEI Jiandong, CHEN Changxi, LI Hongliang
DOI: 10.3969/j.issn.2095-9400.2022.08.009

Objective: To investigate the effect of sodium valproate on mice model of non-alcoholic fatty liver disease (NAFLD) induced by high-fat and high-cholesterol diet. Methods: Forty-two male C57BL/6J mice aged 7-8 weeks were randomly divided as the control group, the model group, and the intervention group. The control group was fed with standard diet, and the model group and the intervention group were fed with high-fat and high-cholesterol diet. Eight weeks after initiation of the experimental diets, the intervention group was given sodium valproate (drinking water with 0.4% sodium valproate) for 8 weeks, while the control group and the model group were given the regular drinking water. Mice were sacrificed after the experiment. The body weight, liver index, serum biochemical indexes, liver histopathological changes, and the relationship with prolyl endopeptidase (PREP) were observed in all mice. Results: Compared with the control group, the body weight, liver index and
epididymal fat index of the model group were significantly increased (P<0.05). Compared with the model group,the intervention group had statistically decreased liver index (P<0.05), but there was no significant difference in body weight or epididymal fat index between the two groups (P>0.05). Serum total cholesterol (TC), low-density lipoprotein (LDL), triglyceride (TG), alanine aminotransferase (ALT), alkaline phosphatase (ALP), fasting blood glucose (FBG), and fasting insulin (Insulin) in the model group were significantly higher than those of the control group (P<0.05); compared with the model group, TC, TG and ALT in the intervention group were significantly decreased (P<0.05), while there was no statistical difference in LDL, AST, ALP, FBG, and Insulin between the two groups (P>0.05). The liver tissue in the model group showed obvious steatosis, inflammation and ballooning.After intervention with sodium valproate, steatosis and inflammation were markedly attenuated, and NAFLD activity score (NAS) was significantly decreased (P<0.05). In liver tissue of the model group, the RQ value of PREP mRNA was (2.02±0.13) and the protein activity of PREP was (473.39±15.41) pmol·mg-1·min-1, both higher than those in the control group (P<0.05). In liver tissue of the intervention group, the RQ value of PREP mRNA was (1.86±0.18), showing no significant difference from that of the model group (P>0.05); however, the protein activity of PREP in the intervention group was (397.41±9.63) pmol·mg-1·min-1, significantly lower than that in the model group (P<0.05). There was no statistical difference in the PREP protein level in liver tissues of three groups (P>0.05). Conclusion: Sodium valproate can attenuate liver steatosis and inflammation in NAFLD mice model by inhibiting PREP activity.
2022 Vol. 52 (8): 657-662 [Abstract] ( 365 ) HTML (1 KB)  PDF (1833 KB)  ( 654 )
663 Clinical significance of APRI and FIB4 in schistosomiasis patients with liver fibrosis
LEI Jianzhu, ZU Deling, JING Qizhi
DOI: 10.3969/j.issn.2095-9400.2022.08.010
Objective: To investigate the characteristics and clinical significance of aspartate aminotransferase to platelet ratio index (APRI) and fibrosis index based on the four factors (FIB4) in schistosomiasis patients with liver fibrosis. Methods: A total of 284 schistosomiasis patients with liver fibrosis in Longyou County People’s Hospital and Quzhou People’s Hospital from January 2019 to June 2021 were selected as the observation group, and 410 healthy subjects as the control group. At the same time, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBiL), platelet count (PLT) were tested, and APRI and FIB4 were calculated. Both groups were examined by B-type ultrasound to test the degree of liver fibrosis. Results: There was evidence to suggest that AST [(61.68±20.25)U/L], ALT [(38.50±13.86)U/L], TBiL [(38.58±18.41)μmol/L],APRI (2.67±1.15), FIB4 (8.51±0.71) in the observation group was respectively higher than those in the control group (P<0.001), with AST [(27.15±6.43)U/L], ALT [(21.21±10.16)U/L], TBiL [(11.23±5.01)μmol/L], APRI (0.49±0.16), FIB4 (0.67±0.03), showing significant difference. The PLT of the observation group was [(64.87±34.49)×109/L], lower than that of the control group [(147.92±48.93)×109/L] (P<0.001). The APRI values of hepatic fibrosis grade I subgroup and hepatic fibrosis grade II subgroup in the observation group were (1.81±0.54), (4.40±2.15) and FIB4 values were (5.38±2.34), (14.75±6.68), respectively (P<0.001). Evidence showed that there was positive correlation between APRI and FIB4 (r=0.757, P<0.001). Conclusion: Combined application of APRI and FIB4 in schistosomiasis patients with liver fibrosis can reflect the status of schistosomiasis liver fibrosis, which is of clinical value.
2022 Vol. 52 (8): 663-666 [Abstract] ( 332 ) HTML (1 KB)  PDF (1363 KB)  ( 561 )
667 Pressure and coping style in caregivers of children with acute lymphoblastic leukemia in remission
Chen Haiyan, WANG Runping, PAN Yan
DOI: 10.3969/j.issn.2095-9400.2022.08.011
Objective: To investigate the status of care pressure and their coping style among parents of children with acute lymphoblastic leukemia in remission. Methods: Using convenience sampling method, we have randomly drawn out 116 parents of children with acute leukemia in remission who were hospitalized in the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University. The questionnaire survey was conducted by using the General Information Questionnaire, Caregiver Strain Index (CSI), and Trait Coping Style Questionnaire (TCSQ) were applied. SPSS19.0 was used for statistical analysis. Results: Of 116 cases, 91 caregivers had pressure (CSI≥7), accounting for 90.5%. From the three dimensions of CSI, caregivers scored the highest in the Work and Financial strains, followed by the Physical and Social strains, and relatively lower in the Psychological strain. Among the 13 items of CSI, “financial pressure” accounted for 95.7%, ranking first; followed by “work affected”, accounting for 91.4%. The coping style score of the caregivers had significant difference from the norm (P<0.05). Positive coping style score of the parents was significantly lower whereas negative coping style score significantly higher than the norm. Conclusion: Parents of children with acute lymphoblastic leukemia in remission have great care pressure and negative coping is predominant. Positive coping can reduce caregiving pressure, and therefore medical staff should guide parents of children with acute lymphoblastic leukemia to adopt positive coping style to reduce their care pressure.
2022 Vol. 52 (8): 667-670,674 [Abstract] ( 420 ) HTML (1 KB)  PDF (1386 KB)  ( 713 )
686 Analysis on Wenzhou residents’ cognition, willingness of body donation and its influencing factors
ZHOU Yujia, HU Qianqian, HE Shiyin, XIANG Shipei, LOU Yanqi, YANG Xindong
DOI: 10.3969/j.issn.2095-9400.2022.08.016
Objective: To understand and analyze the cognition, willingness and influencing factors of cadaver donation among Wenzhou residents, so as to provide reference for the effective development of the related donation work. Methods: A random questionnaire survey was conducted on the residents and the floating population in Wenzhou, and the factors affecting cadaver donation were analyzed by variance test. Results:Data showed that half (50.03%) of the residents had limited knowledge about cadaver donation. The willingness rate of cadaver donation among residents in Wenzhou was only 20.58%, affected by age, gender, education, etc.Constraints of traditional way of thinking (38.44%), distrust of receiving stations (36.92%), too complicated donation procedures (15.07%) and personal beliefs were the reasons for their reluctance to donate bodies.Conclusion: To develop the cause of cadaver donation, we must improve publicity work, perfect donation procedures and protect donors’ rights and interests.
2022 Vol. 52 (8): 686-688,封三 [Abstract] ( 404 ) HTML (1 KB)  PDF (1354 KB)  ( 757 )
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