OBJECTIVE: To prepare Gd-DOTA-E-2 targeted contrast agent and evaluate its relaxation properties, cytotoxicity and targeting. METHODS: E-2 was synthesized by F-moc solid phase method, covalently bonded with chelating agent DOTA, meanwhile ultilizing chelating ligand Gd3+ to prepare MRI contrast agent Gd-DOTA-E-2. The structure was characterized by ESI-MS and NMR, and the properties were evaluated by relaxation properties, MTT and mouse MRI contrast assay. RESULTS: ESI-MS and NMR RESULTS showed that Gd-DOTA-E-2 was synthesized successfully, and its relaxation efficiency(k=9.551 mmol-1•L•s-1) is 1.84 times more efficient than a commercially available contrast agent of Gd-DOTA; MTT RESULTS showed that the toxicity of Gd-DOTA-E-2 cell is lower than that of Gd-DOTA; the experimental RESULTS in MRI angiography showed that Gd-DOTA-E-2 is possessing of targeting and can image clearly. CONCLUSION: Gd-DOTA-E-2 is a kind of MRI contrast agent with good stability, low cytotoxicity and targeted properties in vivo,which is valuable in research and application.

Animals ; Cardiovascular System ; metabolism ; GTPase-Activating Proteins ; metabolism ; Humans ; Nerve Tissue Proteins ; metabolism ; Signal Transduction

Alanine Transaminase ; metabolism ; Animals ; Liver ; drug effects ; Male ; Mice ; Plant Extracts ; pharmacology

Autophagy dysregulation, mitochondrial dynamic abnormality and cell cycle re-entry are implicated in the vulnerable neurons of patients with Alzheimer's disease. This study was designed to testify the association among autophagy, mitochondrial dynamics and cell cycle in dividing neuroblastoma (N2a) cells. The N2a cells were cultured in vitro and treated with different concentrations of 3-methyladenine (3-MA). The cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay. They were randomly divided into control group (cells cultured in normal culture medium) and 3-MA group (cells treated with 10 mmol/L 3-MA). The cell cycle was analyzed in the two groups 3, 6, 12, and 24 h after treatment by flow cytometry. Western blotting was used to evaluate the expression levels of mitofission 1 (Fis1), mitofusin 2 (Mfn2), microtubule-associated protein 1 light chain 3 (LC3), cell cycle-dependent kinase 4 (CDK4) and cdc2. The flow cytometry revealed that the proportion of cells in G(2)/M was significantly increased, and that in G0/G1 was significantly reduced in the 3-MA group as compared with the control group. Western blotting showed that the expression levels of Fis1, LC3, and CDK4 were significantly up-regulated in the 3-MA group at the four indicated time points as compared with the control group. Mfn2 was initially decreased in the 3-MA group, and then significantly increased at 6 h or 12 h. Cdc2 was significantly increased in the 3-MA group at 3 h and 6 h, and then dropped significantly at 12 h and 24 h. Our data indicated that 3-MA-induced suppressed autophagy may interfere with the cell cycle progression and mitochondrial dynamics, and cause cell death. There are interactions among cell cycle, mitochondrial dynamics and autophagy in neurons.

Objective:To investigate the role of blood ammonia in the evaluation of the prognosis of septic patients in the emergency department and to compare its value with mortality in emergency department sepsis (MEDS) score.Methods:A retrospective clinical study was conducted to septic patients who were diagnosed in the Emergency Department of West China Hospital of Sichuan University from June 2017 to May 2018, and met the diagnostic criteria established by the diagnostic criteria of the American College of Chest Physicians/Society of Critical Care Medicine in 2001. The subjects who had other diseases that affected blood ammonia level and were lost to follow-up were excluded. MEDS scores were collected, and the survival status of patients was followed up by telephone. The independent samples t test was used to compare the differences between the two groups, receiver operating characteristic (ROC) curve was used to assess the accuracy of the prediction of sepsis mortality, and the logistic regression model was used to explore the value of the combined use of blood ammonia and MEDS score.Results:Eighty subjects were finally included in the study and divided into the 1-week survival group ( n=52), 1-week death group ( n=28); 4-week survival group ( n=37), 4-week death group ( n=43); 12-week survival group ( n=33), 12-week death group ( n=47); 1-year survival group ( n=32), and 1-year death group ( n=48). There was no statistical difference in the demographic characteristics of subjects between the groups. The average blood ammonia level of all the subjects who died was higher than that of the patients who survived in the same period [(116.57 ± 85.33) μmol/L vs (77.63 ± 35.82) μmol/L, (108.53 ± 73.00) μmol/L vs (71.19 ± 32.53) μmol/L, (106.74 ± 71.59) μmol/L vs (69.21 ± 28.84) μmol/L, (105.77 ± 71.14) μmol/L vs (69.50 ± 29.25) μmol/L, P<0.05]. Based on death after one week, four weeks, twelve weeks and one year, the area under ROC curve (AUC) of blood ammonia was 0.668 (95% CI: 0.542-0.793, P=0.014), 0.706 (95% CI: 0.593-0.819, P=0.002), 0.705 (95% CI: 0.592-0.818, P=0.002), and 0.697 (95% CI: 0.582-0.811, P=0.003), respectively. Compared with the use of blood ammonia, lactic acid or MEDS score alone, the combined use of blood ammonia and MEDS score increased the accuracy of prognosis evaluation in sepstic patients ( P<0.05). Conclusions:Blood ammonia has a high value in predicting the short-term and 1-year prognosis of septic patients in the emergency department. The combined use of blood ammonia and MEDS score can further improve its predictive value.

Objective：To observe the inhibitory effects of pr obucol on nuclear factor-κB （NF-κB）-like activity of vascular smooth mus cle cells(VSMCs). Methods:The effects of probucol and H2O2 on NF-κB-like activity of VSMCs were investigated by electrophoretic mobility shift assay(EMSA). Results:NF-κB activation of VSMCs could be induced by H2O2 or new born calf serum(NCS) for 72 h, the gray-measure of the protein bindings measured through computer scanning was 63.9 and 46.6 respe c tively. NF-κB activity of VSMCs stimulated by H2O2 or NCS might be prevent ed by 100 μmol/L probucol, the inhibitory rate was 37.1%, 14.8% respectively. Conclusion:probucol can inhibit NF-κB activity of VSMCs stim ulated by NCS or H2O2. These effects of probucol might help to re duce development of atheroclerosis and restenosis after percutaneous transluminal coronary angioplasty.

Objective To investigate the current prevailing status of endemic fluorosis in the Yellow River basin of Shandong Province and to provide the scientific evidence for making strategies in prevention and control.Methods Nine counties were chosen to carry out the epidemiological investigation.The content of fluoride in drinking water was determined by F-ion selective electrode and dental fluorosis of children aged 8～12 years old was diagnosed by Deans method.Results Water fluoride content was determined in 1761 fluorosis villages,among which 606 villages had water fluoride content≤1.00 mg/L,accounting for 34.41%(606/1761);1155 villages had water fluoride content＞1.00 mg/L,which accounted for 65.59%(1155/1761).The highest water fluoride content was 11.33 mg/L.Water fluoride content of 618 water-improving and defluoridation projects had been determined,among which 449 projects had water fluoride content≤1.00 mg/L and accounted for 72.65%(449/618),169 projects had water fluoride content＞1.00 mg/L and accounted for 27.35%(169/618),the highest water fluoride content was 5.85 mg/L.The total rate of dental fluorosis of children aged 8～12 years old was 45.03%(25 579/56 804) and the index of dental fluorosis was 0.80. Conclusions In the Yellow River basin in Shandong Province,up to 50.00%in the villages the water fluoride content exceeds the county standard(≤1.00 mg/L).The prevalence of endemic fluorosis in the basin hasn't been effectively controlled.So the counterrneasures for endemic fluorosis should be carried out as soon as possible.

OBJECTIVETo study mRNA and protein expression of HOXA5 gene in breast carcinoma, to correlate the expression of HOXA5 gene with clinicopathologic parameters and to explore the possible role of HOXA5 gene in carcinogenesis, progression and metastasis of breast carcinoma.

METHODSTaqMan real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was applied on 60 cases of primary breast carcinoma and 24 cases of benign mammary lesions in order to detect mRNA expression of HOXA5 gene. Immunohistochemical study using polyclonal antibody against HOXA5 was also performed. Statistical analysis was carried out to analyze the correlation between HOXA5 gene expression and various clinical parameters in these breast cancer patients.

RESULTS(1) The relative expression level of HOXA5 mRNA ranged from 0.73 to 193.07 (average = 20.85) in primary breast carcinoma, in contrast to 5.42 to 81.91 (average = 30.94) in benign mammary lesions. Compared with benign mammary lesions, a significant reduction in expression of HOXA5 mRNA was noted in primary breast carcinoma (P < 0.01). (2) There was a decreased or completely diminished HOXA5 protein expression in breast carcinoma. (3) HOXA5 mRNA expression was significantly lower in lymph node-positive cases, when compared with that in lymph node-negative cases (P < 0.05). A significant difference of HOXA5 protein expression was also observed in both groups (P < 0.01). Immunohistochemical staining of HOXA5 was either negative or weakly positive in lymph node-positive cases. On the other hand, moderately or strongly positive HOXA5 staining was noted in lymph node-negative cases. (4) Neither mRNA nor protein expression of HOXA5 gene correlated with clinicopathologic parameters such as age of patients, size of tumor, clinical stage, pathologic subtype or histologic grade (P > 0.05).

CONCLUSIONSDisordered expression of HOXA5 gene may play a role in the carcinogenesis of breast cancer. Reduced expression of HOXA5 gene may be related to the metastatic potential of breast carcinoma cells.

Adult ; Aged ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Homeodomain Proteins ; biosynthesis ; genetics ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; Papilloma, Intraductal ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

BACKGROUNDApoptosis is a major cause of ischemic heart dysfunction. Apelin, the endogenous ligand for the G-protein-coupled APJ receptor, has been reported to exert cardioprotective effects during myocardial injury. The aim of this study was to investigate the effects of apelin on apoptosis of rat cardiomyocytes induced by glucose deprivation (GD) and study the related signaling pathway.

METHODSApelin and APJ mRNA expression were determined by RT-PCR in neonatal rat cardiomyocytes during different durations of GD. Cardiomyocyte apoptosis was detected by annexin V-FITC/propidium iodide (PI) staining after GD for 12 hours with or without apelin-13 (10 and 100 nmol/L) pretreatment. Protein levels of Akt and the mammalian target of rapamycin (mTOR) as well as cell apoptosis were detected in the presence or absence of LY294002 (a phosphatidylinositol 3-kinases (PI3K) inhibitor) or rapamycin (a mTOR inhibitor).

RESULTSApelin mRNA expression was up-regulated when cardiomyocytes were exposed to GD for 6, 12, 18, and 24 hours compared with the base level (P > 0.05, P < 0.01, P < 0.01, P < 0.01). However, when cardiomyocytes were exposed to GD for up to 36 hours, apelin mRNA expression was 17% lower than the base level (P < 0.05). APJ mRNA expression paralleled that of apelin. Apelin-13 pretreatment at 100 nmol/L significantly inhibited GD-induced cardiomyocyte apoptosis (P < 0.05) and increased Akt and mTOR phosphorylation (P < 0.01, P < 0.01). At the same time apelin-13 (100 nmol/L) up-regulated Bcl-2 protein expression and down-regulated Bax and cleaved caspase-3 expression (P < 0.01, P < 0.05, P < 0.05). The anti-apoptotic effect of apelin-13 was blocked by LY294002 (P < 0.01) but not by rapamycin.

CONCLUSIONSThe endogenous apelin-APJ system is compensatorily up-regulated and ultimately down-regulated following sustained myocardial ischemia. Apelin protects against ischemic cardiomyocyte apoptosis via activation of the PI3K/Akt pathway.

Animals ; Apelin ; Apelin Receptors ; Apoptosis ; Carrier Proteins ; genetics ; physiology ; Caspase 3 ; analysis ; Cell Survival ; Cells, Cultured ; Glucose ; deficiency ; Intercellular Signaling Peptides and Proteins ; Myocytes, Cardiac ; physiology ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; physiology ; Signal Transduction ; bcl-2-Associated X Protein ; analysis

OBJECTIVETo investigate the dynamic changes of the epididymal size 1 year after vasectomy.

METHODSFifty male volunteers received vasoligation. Before and at 1, 2, 3, 6, and 12 months after operation, we measured the size and detected the internal echoes of the epididymis using color Doppler ultrasonography.

RESULTSThe bilateral epididymides were both thickened post-operatively in all the 50 cases, with statistically significant differences between the baseline and the 1st month, the 1st and the 2nd month, the 2nd and the 3rd month, or the 3rd and the 6th month after surgery (all P < 0.01), but not between the 6th and the 12th month (P > 0.05).

CONCLUSIONWithin 6 months after vasectomy, the bilateral epididymides manifested a progressive thickening, but basically restored their balance of secretion-absorption after 6 months.

Epididymis ; diagnostic imaging ; pathology ; physiology ; Humans ; Male ; Organ Size ; Postoperative Period ; Time Factors ; Ultrasonography, Doppler, Color ; Vasectomy