Angiolymphoid Hyperplasia with Eosinophilia ; pathology ; Forehead ; pathology ; Humans ; Male ; Middle Aged ; Parotid Gland ; pathology

Objective:To investigate mechanisms underlying the signal crosstalk of VEGF-IL-6-STAT3 between cutaneous melanoma cells and vascular endothelial cells.Methods:EC-304 vascular endothelial cells were divided into 3 groups: control group cultured in conventional endothelial cell-conditioned medium, vascular endothelial growth factor (VEGF) group cultured in endothelial cell-conditioned medium containing 50 μg/L VEGF 165, A375 co-culture group co-cultured with a melanoma cell line A375. After 24-, 48- and 72-hour treatment, the culture medium was collected, and enzyme-linked immunosorbent assay was performed to detect the level of interleukin-6 (IL-6) . Cultured A375 cells were divided into 4 groups: control group receiving conventional culture in Dulbecco′s modified Eagle′s medium (DMEM) , A375+ EC-304 group co-cultured with EC-304 cells, A375+ EC-304+ IL-6 group co-cultured with EC-304 cells in DMEM containing 50 μg/L IL-6 (an agonist of the signal transducer and activator of transcription-3 [STAT3] pathway) , A375+ EC-304+ JSI-124 group co-cultured with EC-304 cells in DMEM containing 1 μmol/L JSI-124 (a STAT3 pathway inhibitor) . After 24-, 48- and 72-hour treatment, cells were collected, and Western blot analysis, cell counting kit-8 (CCK8) assay and Transwell invasion assay were performed to determine the protein expression of STAT3 and phosphorylated (p) -STAT3, cellular proliferative activity and invasive activity, respectively. Two-way analysis of variance and t test were used for statistical analysis. Results:The level of IL-6 significantly increased in the culture medium of EC-304 cells in the VEGF group and A375 co-culture group compared with the control group ( FVEGF = 29.63, P < 0.001; FA375 = 11.09, P = 0.020) . Compared with the control group, the A375+ EC-304 group showed significantly enhanced protein expression of p-STAT3 in A375 cells ( P < 0.001) , increased cell activity ( P < 0.001) , and increased number of invasive cells (152.66 ± 16.04 vs. 86.13 ± 7.24, t= 4.43, P < 0.001) ; compared with the A375+ EC-304 group, the A375+ EC-304+ IL-6 group showed significantly increased protein expression of p-STAT3 ( P < 0.001) , enhanced cell activity ( P < 0.001) , and increased number of invasive cells (187.34 ± 14.38, t= 2.17, P < 0.001) ; compared with the A375+ EC-304 group, the A375+ EC-304+ JSI-124 group showed significantly decreased protein expression of p-STAT3 ( P < 0.001) , decreased cell activity ( P < 0.001) , and decreased number of invasive cells (124.92 ± 8.72, t=-1.86, P < 0.001) . Conclusion:There is a signal crosstalk of VEGF-IL-6-STAT3 between cutaneous melanoma cells and vascular endothelial cells, which may play an important role in the proliferation and invasion of A375 cells.

Objective To improve the method to establish a rat model of vascular dementia and to better serve the experimental studies on vascular dementia.Methods We used the method of“repeatedly clipping the carotid artery com-bined with injection of sodium nitroprusside and with permanent unilateral carotid artery ligation” to prepare a rat model of vascular dementia.The drug piracetam was used to validate the established rat model in respect of the behavior and histopa-thology.Results Different from the reports of previous research, firstly, the results of this study suggested that injection dose of sodium nitroprusside should be 2.0 mg/kg, room temperature should be controlled at 28℃ during surgical opera-tion, and kept at 25℃postoperatively for 24 hours.Under these experimental conditions, the rats were stable and the death rate was reduced.Secondly,“repeatedly clipping carotid artery combined with permanent unilateral carotid artery ligation”could cut off about a third of cerebral blood supply, and causing chronic cerebral ischemia, which is seemingly more con-sistent with the pathogenesis of vascular dementia.Experimental results showed that compared with the control group, the navigation incubation period was extended and space search ability became worse in the model group, cell number was de-creased, with blurred cell contour and deeply stained cytoplasm, and cell nuclei were not clear in the hippocampal tissue. Conclusions Our findings indicate that the improved methodrepeatedly clipping the carotid artery combined with injec-tion of sodium nitroprusside and with permanent unilateral carotid artery ligationcan be used to efficiently establish a rat model of vascular dementia.The similar results of experiment using piracetam validate that this rat model can be used in vascular dementia-related experimental studies.

AIM To establish the methods for assaying the activities of tolbutamide hydroxylase (CYP2C8/9) and chlorzoxazone 6-hydroxylase(CYP2E1) from adult human livers microsomes. METHODS The microsomes was isolated from human adults liver and the contents of microsomal protein were determined. Using the tolbutamide and chlorzoxazone as substrates, the amount of hydroxytoblbutamide and hydroxychlorzoxazone formed during the incubation period was quantified by extroppolating from the standard curve and the specific activity of CYP2C8/9 and CYP2E1 were calculated. RESULTS There was no siginificant influence on the activities of CYP2C8/9 and CYP2E1 in different times. CONCLUSIONS The methods utilized to estimate the activity of CYP2C8/9 and CYP2E1 were simple, stable, and repeatable. These methods can be used in new drug screening, safety evaluation and reseasch on pathology and toxicology of liver.

Objective To evaluate the effectiveness of mindfulness-based stress reduction( MBSR) on anxiety and depression in spouses of breast cancer patients and to analyze the mediating effect of caregiver burden for the training effects. Methods Randomized control trial was used in this research.100 spouses of breast cancer patients were recruited and they were randomized into experimental group or control group. 91 of them completed the study finally (45 in experimental group and 46 in control group). Participants in con-trol group received no intervention,while participants in experimental group received MBSR once a week for 8 weeks. All participants were assessed with the Zarit Caregiver Burden Interview ( ZBI,measuring caregiver burden) ,Beck Depression Inventory( BDI,measuring depression level) and Hamilton Anxiety Scale( HAMA, measuring anxiety level) before and after intervention. Results After 8-week intervention,participants in experimental group got significantly lower total scores of ZBI,BDI and HAMA compared with pretest ((22.31 ±5.76) vs (25.98±6.00),(8.47±3.21) vs (11.87±3.51),(14.98±4.75) vs (17.93±4.65), P<0. 01), while there were no significant differences between posttest and pretest on any score in control group (P>0.05).Caregiver burden played mediating effect in MBSR for improving depression,and mediating effect was 56.6% of the total effect. Conclusion MBSR can effectively improve anxiety and depression with the spou-ses of breast cancer patients,and the caregiver burden is the intermediary variable in the effect of MBSR for improving depression.

Objective To study the expression ot secreted proteins in aggregated dermal papilla cells (DPCs).Methods DPCs were isolated from human scalp tissue and subjected to primary culture and subculture.Aggregated and non-aggregated DPCs served as the subject of this study.Secreted proteins were prepared from these cells and subjected to two-dimensional polyacrylamide gel electrophoresis.Differentially expressed proteins were screened by the PDQuest image analysis software.Protein spots were digested and identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry,and finally analyzed using the National Center for Biotechnology Information (NCBI) non-redundant (Nr) protein database.Results Two-dimensional electrophoresis maps with good repeatability and high resolution were established.Image analysis of 2-D gels revealed that the average number of detected protein spots was 1 134 ± 52 and 1 078 ± 36 in aggregated and nonaggregated DPCs respectively,and the majority of these protein spots were matched between aggregated and nonaggregated DPCs.Twenty-eight protein spots showed more than 5-fold difference between the two groups of cells,and 10 proteins were preliminarily identified as differentially expressed proteins by peptide-mass fingerprinting.Of these differentially expressed proteins,8 proteins including Rhogdi 1,filamin A,cystatin C,fibronectin,cyclophilin A,procollagen C proteinase enhancer 1,tissue inhibitor of metalloproteinase and tissue inhibitor of metalloproteinase-2 were up-regulated,and 2 proteins including neuropolypeptide h3 and matrix metalloproteinase-3/tissue inhibitor of metalloproteinase-1 complex were down-regulated in aggregated DPCs compared with non-aggregated DPCs.Conclusions Differentially expressed proteins between aggregated and non-aggregated DPCs are mainly implicated in cell signaling pathway,cellular proliferation and differentiation,extracellular matrix synthesis and degradation,and so on.

With the advance of drug development and research techniques, the drug metabolic processes and mechanism can be more deeply achieved. As the drug metabolism and pharmacokinetics process are mediated by drug metabolizing enzymes and transporters, study of drug metabolizing enzymes and transporters has become an important part for drug development. The traditional immunoassays with low sensitivity and poor specificity can not reflect the accurate expression level of drug metabolizing enzymes and transporters. We now give a brief review on the quantitative study of drug metabolizing enzymes and transporters by mass spectrometry-based proteomic approach.

Previous studies have revealed Twist,a basic helix-loop-helix transcription factor,plays an important role in breast cancer metastasis.To clarify the molecular mechanism of its involvement in cancer metastasis,Twist was silenced by RNAi in highly metastatic 4T1 cells.Then microarray chips were used to investigate the gene-expression pattern of the Twist-knockdown 4T1 cells and the normal 4T1 cells.The results indicated that silencing of Twist significantly suppressesed lung metastasis of 4T1 cells in vivo.Direct comparison of gene-expression profiles showed that 167 genes in Twist-knockdown cells differed dramatically in expression levels from those in control cells.Among the 167 genes,26 well-known tumor-associated genes,including 15 up-regulated and 11 down-regulated genes were found.These genes appear to be regulated by Twist during breast tumorigenesis.The findings provide new insights into the mechanism by which Twist is involved in tumorigenesis.

Objective To retrospectively evaluate the mammographic features of breast ductal carcinoma in situ (DCIS)and DCIS with small invasive foci,and to analyze the correlation between the mammographic findings and the prognostic biologic factors.Methods The mammographic examination was performed in 95 consecutive women with breast DCIS(n = 50)and DCIS with invasive foci(n = 45 ).The prognostic biologic factors including progesterone receptor(PR),C-erbB-2,and p53 were evaluated in 62 of 95 cases.Categorical data were expressed as percentages and analyzed by using the X~2 test,and furthermore the odds ratio was measured.Results(1)Only one abnormality was seen on mammography in 62 patients. Combined two abnormalities on mammography were seen in 26 patients.Mammograms were normal in 7 patients.(2)Calcifications with or without other abnormality were noted in 62 cases.Of them,73% (n =45)had higher probability of malignancy calcifications and the others were intermediate concern calcifications.Clustered calcifications(36 lesions)was the most common distribution,which usually accompanied by another abnormality.And then were segmental(18 lesions)distributed pattern.As far as the shape of mass (n = 22)was concerned,the oval shaped lesion(13 cases)was the most common,and the margin of the mass appeared as ill-defined in 15 eases,microlobulated in 1,circumscribed in 4,and obscured in 2,respectively.Isodensity mass had a higher frequency in this group(12/22,55%).Other non-calcification findings included architecture distortion(7 cases),local asymmetry (15 cases),global asymmetry (5 cases),and solitary dilated duct (3 cases),and most of them accompanied with other signs. (3)For expression profile of the biological factors,significant differences were found among malignant calcification group,intermediate concern calcification group,and non-calcification group. The odds of PR positive for the lesions noted as non-calcification were 11.00 times higher (X~2 =8.571 ,P=0.003 ;95% CI, 1.998—60.572)than the lesions noted as intermediate concern calcifications,and 8.80 times higher (X~2 = 9.748,P=0.002 ;95% CI,2.024—38.253)than the lesions noted as malignant calcifications.The odds of C-erbB-2 positive for the lesions showed as malignant calcifications were 12.35 times higher (X~2=7.353, P=0.007 ;95% CI,1.447—105.443)than the lesions showed as non-calcification,and 5.74 times higher (X~2=4.977,P = 0.026;95% CI,1.110—29.645)than the lesions showed as intermediate concern calcifications.Conclusion The mammographic features of DCIS and DCIS with small invasive foci were characteristic.Mammographic findings could be a prognostic markers,which could provide a possibility for making a treatment plan.

With the advance of drug development and research techniques, the drug metabolic processes and mechanism can be more deeply achieved. As the drug metabolism and pharmacokinetics process are mediated by drug metabolizing enzymes and transporters, study of drug metabolizing enzymes and transporters has become an important part for drug development. The traditional immunoassays with low sensitivity and poor specificity can not reflect the accurate expression level of drug metabolizing enzymes and transporters. We now give a brief review on the quantitative study of drug metabolizing enzymes and transporters by mass spectrometry-based proteomic approach.
Enzymes ; chemistry ; Humans ; Inactivation, Metabolic ; Mass Spectrometry ; Membrane Transport Proteins ; chemistry ; Pharmacokinetics ; Proteomics