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Objective:To observe the role of neuroinflammation in cognitive dysfunction induced by 1-bromopropane (1-BP) in rats.Methods:Male Wistar rats were randomly divided into control group, 1-BP group, pyrrolidine dithiocarbamate(PDTC)+ 1-BP group and PDTC group, with 15 rats in each group. Rats in 1-BP group and PDTC+ 1-BP group were given 800 mg / kg 1-BP by gavage, and rats in control group and PDTC group were given equal volume corn oil once a day for 12 days; rats in PDTC group and PDTC+ 1-BP group were intraperitoneally injected with 100 mg / kg PDTC 30 minutes after gavage, while rats in control group and 1-BP group were injected with equal volume of normal saline once a day for 12 days.From the 7th to 12th day of the experiment, ten rats in each group were randomly selected and subjected to Morris water maze test for detect the cognitive function. In the positioning navigation test, the learning ability of rats was evaluated by the escape latency and total swimming distance respectively. In the space exploration experiment, the memory ability of experimental animals was evaluated by the number of times crossing the target platform. After the experiment, ten rats were sacrificed, the cerebral prefrontal cortex was harvested. The cytosolic and nuclear NF-κB expression and phosphorylation were detected by Western blot, the mRNA levels of TNF-α and IL-1β were detected by qRT-PCR. After cardiac perfusion fixation, the brains of 5 rats were taken to make frozen sections for immunohistochemical staining and Nissl staining. SPSS 20.0 software was used for statistical analysis, repetitive measurement deviation analysis was used for the analysis of the swimming distance and the escape latency in positioning navigation test, One-way ANOVA was used for the analysis of the number of times crossed the target platform in spatial probe test and other data. Tukey's test was used for Post hoc comparison.Results:The results of Morris water maze showed that there was significant interaction between group and training time in the total swimming distance of rats in the four groups ( F=3.762, P<0.05). Simple effect analysis showed that the total swimming distance of 1-BP group in 1-4 days were longer than those of control group (all P<0.05), while the total swimming distance of PDTC+ 1-BP group in 1-4 days were shorter than those of 1-BP group (all P<0.05). There was significant interaction between group and training time in the escape latency among the four groups ( F=6.541, P<0.01). The escape latencies of 1-BP group in 1-4 days were longer than those of control group (all P<0.05), while the escape latencies of PDTC+ 1-BP group in 1-4 days were shorter than those of 1-BP group (all P<0.05). The results of space exploration experiment showed that there was significant difference in the number of crossing the platform among the four groups ( F=75.333, P<0.01). The number of crossing the platform (1.08±0.29) in 1-BP group was lower than that in the control (3.35±0.05) ( P<0.01). The number of crossing the platform (1.95±0.26) in PDTC+ 1-BP group was higher than that in 1-BP group ( P<0.01). It had significant difference both in the cytoplasm and in the nucleus of the NF-κB protein level in prefrontal cortex among rats of the four groups ( F=20.865, 23.877, both P<0.01). The levels of NF-κB in cytoplasm and nucleus of rats in 1-BP group were both higher than those in control group (cytoplasm: (177.3±32.1)%, (100.0±8.4)%, P<0.01; nucleus: ( 173.2±27.1)%, (100.0±8.4)%, P<0.01). While the levels of NF-κB in cytoplasm and nucleus of 1-BP+ PDTC group were both lower than those of 1-BP group (cytoplasm: (148.7±22.0)%, (177.3±32.1)%, P<0.01; nucleus: (149.7±18.8)%, (173.2±27.1)%, P<0.01). The results of qRT-PCR showed that there were significant differences in the mRNA levels of TNF-α and IL-1β in the prefrontal cortex among the four groups ( F=17.464, 17.382, both P<0.01). The levels of TNF-α and IL-1β mRNA in 1-BP group were higher than those in control group (both P<0.05), and the levels of TNF-α and IL-1β mRNA in PDTC+ 1-BP group were both lower than those in 1-BP group (both P<0.05). The results of immunohistochemistry showed that compared with the control group, the number of microglia and astrocytes in the 1-BP group increased (microglia: (158.30±9.68), (110.20±16.30), P<0.05; astrocytes: (122.76±4.35), (80.24±6.96), P<0.05), and the morphology was also activated, with light staining and reduced number of Nissl bodies in neurons.The number of microglia and astrocytes in PDTC + 1-BP group was lower than that in 1-BP group (microglia: (131.70±14.67), (158.30±9.68), P<0.05; astrocytes: (101.54±4.55), (122.76±4.35), P<0.05), and the Nissl body staining of neurons was significantly deepened. Conclusion:NF-κB signaling pathway might be the key mechanism of 1-BP neurotoxicity. PDTC intervention could significantly improve the neuroinflammatory response and behavioral disorders of experimental animals intoxicated with 1-BP.
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Objective:To explore the factors influencing patient safety behavior of nursing students during clinical practice.Methods:A cross-sectional study was used, in which a total of 214 participants completed the online survey related to the undergraduate nursing students ′ clinical teaching satisfaction scale, safety attitude questionnaire and nurse safety behavior questionnaire. Results:The mean scores of clinical teaching satisfaction, safety attitude and nurse safety behavior were (3.83±0.81), (3.70±0.65) and (3.93±0.79) respectively. There were positive correlations between clinical teaching satisfaction, safety attitude with the nurse safety behavior ( r=0.82, 0.75, P<0.01). The results also showed that the same shifts with their clinical instructors, clinical teaching satisfaction and safety attitude could affect safety behavior of nursing students ( F=67.81, R2=0.753, ΔR2= 0.742, P=0.00), which accounted for 74.20% of total variation. Conclusions:The same shift arrangement for both clinical instructors and nursing students, enhancement of the demonstration and guidance from clinical instructors, as well as strengthening the cultivating patient safety attitude would be beneficial to promoting the safety behavior of nursing students during clinical practice.
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Objective To study the effect of estrogen on proliferation of astrocytes in hippocampus of mice following middle cerebral artery occlusion(MCAO).Methods One hundred and eight Kunming mice were randomly divided into estrogen group(n=54)and saline group(n=54).The animals in two groups underwent right MCAO with tissue samples taken at 3,6,12,24,48and 72h after MCAO.The ischemic site was detected and the ischemic size was measured with TTC staining,the damage of neurons in hippocampus was assayed with HE staining,the expression of GFAP in hippocampal astrocytes was detected with immunohistochemical staining.Results The cerebral infarction size was significantly smaller in estrogen group than in saline group at different time points after MCAO(P＜0.05,P＜0.01)especially at 12hafter MCAO(31.50％±3.36％vs 54.50％±5.68％,P=0.019).The damage of hippocampal neurons aggregated with the prolonged ischemia time in two groups and was milder in estrogen group than in saline group at the same time points.The expression level of GFAP positive cells in bilateral hippocampal areas was higher when the ischemia time was prolonged and was significantly higher in ischemic hippocampus of estrogen group than in that of control group except at 6hin CA3ischemic area(P＜0.05).Conclusion Estrogen can protect mice against focal cerebral ischemia,stimulate the genesis of astrocyte synapses,alleviate neuronal damage after ischemia,and can thus reduce the size of cerebral infarction.
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Objective: To determine the expression of TAZ and its role in angiogenesis in gastric carcinoma. Methods: Immunohistochemical staining was performed to investigate the expression of TAZ and to determine whether a direct relationship exists between TAZ and β-catenin. Transfection with TAZ overexpression plasmid in MKN28 cells was conducted to induce exogenous expression of TAZ and a TAZ knockdown plasmid was transfected into MGC803 cells to reduce TAZ levels. The effects on endothelial cell formation, proliferation, and migration were determined by Matrigel three-dimensional culture, MTT proliferation assay and Transwell migration assay. In addition, the expression of TAZ and β-catenin in transfected gastric cancer cells was detected by Western blot. Results: Immunohistochemistry showed that TAZ protein was expressed in 64 of 150 gastric cancer sample tissues (43%), TAZ was localized in the nucleus, and its expression was associated with tumor grade, TNM stage, metastasis, and microvessel density (MVD) (P<0.05). In addition, the expression frequency of β-catenin in the TAZ positive group was 67.2%, which was significantly higher than that in the TAZ negative group, and the expression of TAZ was positively correlated with β-catenin. After transfection, TAZ overexpression increased the expression of β-catenin and enhanced HUVECs tube formation, proliferation, and migration. In the MGC803 cells transfected with the knockdown plasmid, β-catenin levels were decreased and HUVECs motility was inhibited. Conclusions: TAZ may promote angiogenesis in gastric cancer by promoting β-catenin expression.
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Objective:To explore the diagnostic value of ultrasound-guided fine needle aspiration biopsy(US-FNAB)for thyroid nod-ules.Methods:The clinical characteristics and cytopathological diagnosis of patients with thyroid nodules in Tianjin Medical University General Hospital were analyzed retrospectively;the results of the cytopathological and pathological diagnoses were compared and an-alyzed.Results:Of the 1,241 US-FNAB samples,the ratio of men to women with thyroid nodules was 1:3.83(257/984).The incidence of thyroid nodules gradually increased from the age of 20 years and declined after the age of 60 years.The nodules,which were less than or equal to 1.0 cm in size,accounted for 51.57%(640 cases),and Thyroid Imaging Reporting And Data System(TIRADS)classifica-tion 4 accounted for 86.38%(1 072 cases).Of cyto-pathological diagnoses,22.00%(273 cases)were non-diagnostic,9.75%(121 cases) were benign,30.62%(380 cases)were atypia with undetermined significance,32.15%(399 cases)were suspicious for malignancy,and 5.48%(68 cases)were malignant.In the 302 patients who underwent surgery,the number of cases of clear diagnoses,unavailable di-agnoses,and atypia of undetermined significance were 203,21,and 78,respectively.In the 203 cases of clear diagnoses,the sensitivi-ty,specificity,positive predictive value,negative predictive value,precision,and misdiagnoses following US-FNAB of thyroid nodules were 100.00%(201/201),50.00%(1/2),99.50%(201/202),100.00%(1/1),99.51%(202/203),and 0.49%(1/203),respectively.In the 78 cases that were atypia of undetermined significance,the malignancy rate was 70.51%.Whether the atypia of undetermined signifi-cance was malignant or not was related to the TIRADS classification(P<0.05),and not related to the age,sex,tumor size,or location of the nodules(P>0.05).Conclusions:US-FNAB has high diagnostic value for thyroid nodules and is worthy of being popularized widely.If it replaced some intra-operative frozen sectioning procedures,it may reduce intra-operative waiting time and financial burden of pa-tients.
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Objective:To examine the expression of IQGAP1 in hepatocellular carcinoma and its effect on vasculogenic mimicry(VM). Methods:Immunohistochemical staining was performed to investigate the expression of IQGAP1.CD31/PAS double staining was per-formed to detect VM and analyze the correlation of IQGAP1 and VM.HepG2 cells were transfected with an IQGAP1 overexpression plasmid to induce exogenous expression of IQGAP1,and an IQGAP1 knockdown plasmid was transfected into SMMC7721 cells to re-duce IQGAP1 levels.The expression of cancer stem cell markers CD133,CD44,Sox2,and ALDH1 was analyzed by Western blot and compared with that in the control.Cellular functional experiments were used to determine the role of IQGAP1 in promoting cancer cells' ability of invasiveness and migration, proliferation, and VM formation. Results: Immunohistochemical analysis revealed that IQGAP1 was mainly located in the cell membrane and/or cytoplasm,and the staining intensity was correlated with tumor grade,me-tastasis,and VM(P<0.05).Cells transfected with the overexpression plasmid showed enhanced CD133,CD44,Sox2,and ALDH1 levels due to the increase in IQGAP1 and exhibited increased invasion ability,proliferation,and VM formation.In the SMMC7721 cells trans-fected with the knockdown plasmid,CD133,CD44,Sox2,and ALDH1 levels were decreased and motility was inhibited.Conclusions:IQGAP1 supports malignant behavior in hepatocellular carcinoma and may promote VM by increasing stemness.
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Objective@#To evaluate and compare the caries prevention effectiveness of a resin-based sealant and a glass ionomer sealant on permanent first molars in 7- to 9-years-old school children.@*Methods@#A randomized controlled, examiner-blinded, 5-year clinical trial was conducted in 19 primary schools in Dalian, Liaoning province. Totally 419 children (with 664 first molars) who had at least one sound permanent first molar with deep fissures or fissures with signs of non-cavitated lesions were recruited and stratified and allocated randomly into three parallel groups. A resin-based sealant was used in Group R (136 children with 219 first molars) and a glass ionomer sealant was used in Group G (130 children with 218 first molars). The third group (Group N, 153 children with 227 first molars) with none of the two sealants applied was served as blank control. Follow-up examinations were conducted at 0.5, 1, 2, and 5 years after sealing in order to assess the retention of the sealants and the incidence of caries occurrence. Chi-square test was adopted to compare the differences of caries incidence among the 3 groups. Generalized estimating equations (GEE) was used to analyze the effective factors of caries incidence.@*Results@#During the 5-year follow-up period, 13.4% (23/172) of the teeth in Group R, 22.5% (40/178) of the teeth in Group G and 34.5% (57/165) of the teeth in Group N were diagnosed as having developed cavitated lesions. And the differences in the incidence of caries among the three groups was statistically significant (P<0.05). According to GEE, the resin-based sealant and the glass ionomer sealant were found to be the protection factors and the non-cavitated lesion before sealing was found to be the risk factor. The resin-based sealant was found to be the protection factor to the glass ionomer sealant when the blank control group was removed.@*Conclusions@#Both resin-based sealant and glass ionomer sealant could effectively prevent the permanent first molars from the occurrence of carious lesions in 5 years. The resin-based sealant was superior to the glass ionomer sealant in caries prevention.
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Objective:To investigate the prevalence and related factors of dental caries in 3-year-old children in the Development District in Dalian.Methods:A multi-stage,randomized,cluster sampling method was used in the cross-sectional study.All 3-year-old children in the selected 15 kindergartens were invited to take part in the oral health examination conducted by 3 calibrated examiners.The WHO recommended caries diagnosis criteria were used.The structured questionnaire including the information of children's oral hygiene habits,dietary habits,parents' oral health knowledge and socioeconomic status was completed by the parents.Results:1 220 3-year-old children completed the clinical oral examination and 964 children's parents(79.0％) finished the questionnaire.The caries prevalence in 3-year-old children was 62.8％,and the mean dmft and dmfs scores were 3.40 and 7.34,respectively.No differences in the prevalence of caries,mean dmft and dmfs scores were found between boys and girls(P ＞0.05).Logistic regression analysis showed that the frequency of sweets and sugar-containing beverages consumption(P＜0.05),parents'education level(P＜0.05) were significantly related to the caries prevalence.Conclusion:The prevalence and severity of dental caries in 3-year-old children in Dalian Development District were high and the frequency of consumption sugar-containing foods and beverages,parents'education level were related to the decay prevalence.
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OBJECTIVE To observe the neurotoxicity of 1-bromopropane(BP) and investigate the protective effects of edaravone(Edv) against BP-induced deficits of spatial learning and memory ability in rats by its anti-inflammatory mechanism. METHODS Adult male Wistar rats were ig given BP 800 mg·kg-1 to develop the model, followed by Edv 1, 3 and 5 mg·kg-1 ip treatment respectively 4 h later for consecutive 12 d. From the 7th day (d 7), all rats were subjected to the five-day place navigation in Morris water maze (MWM) to measure the escape latency and the total swimming distance. On d 6 of MWM, spatial probe test was performed and the crossing times of rats were recorded to evaluate the spatial memory ability. At the end of the behavioral experiment, four rats in each group were randomly selected and the frozen section of the whole brain was sliced for thionin staining and immunohisto?chemistry. The other eight sacrifced rat brains from each group were harvested for the determination of the tumor necrosis factor-α (TNF-α) and nitric oxide (NO) by ELISA and nitrate reductase method, respectively. RESULTS The results of MWM test showed that compared with control rats the escape latencies of rats in BP group were increased by 60.8%, 81.9%,124.0% and 323.3%, respectively, during the d 2-d 5 of MWM, and the total swimming distance increased by 47.0%, 66.4%, 106.0% and 277.6%, respectirely. All the differences between BP group and control group were significant (P<0.05, P<0.01). In the spatial probe trial, the crossing times of rats in BP group were significantly decreased, compared with the control rats (P<0.01). Morphologically, thionin staining and immunohistochemistry revealed significant microglia activation and neuron loss in the rat forebrains, accompanied by a 147.6% and 18.7% increase in NO and TNF-α levels in rats treated with BP respectively compared with control values (P<0.05, P<0.01). After co-treatment at different dosages of Edv with BP, the escape latencies of rats in BP+Edv 5 mg·kg-1 group were decreased by 38.4%and 44.3%(P<0.01), and the total swimming distance decreased 34.5%and 43.3%(P<0.05, P<0.01), respectively, compared with the BP treated rats on the d 4 and d 5 of MWM test. The microglia activation and neuron damage in the brain of rats induced by BP treatment were significantly alleviated in BP+Edv groups. In addition, the contents of NO and TNF-α were decreased in BP+Edv 1, 3 and 5 mg · kg-1 groups, with a decrease of 53.8%, 55.4% and 59.8% in NO, and 12.2%, 15.8% and 22.2% in TNF-α(P<0.05, P<0.01), respectively. CONCLUSION Edv could effectively protect against central neurotoxicity induced by BP via anti-neuro?inflammation.
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Objective:This study aims to determine the suitable cell line to be used in isolating cancer stem cells by comparing the characteristics of tumor stem cells in renal cell carcinoma cell lines SN12C and 786-O. Methods:The rate of sphere formation in SN12C and 786-O cells was determined in serum-free medium (SFM). The expression levels of CD133, CD44, Nanog, and Oct3/4 were investi-gated through flow cytometry. Moreover, the tumorigenicity of spheroid cell that originated from SN12C and 786-O cells was investi-gated in vivo by using a tumor model. Results:The average time of sphere formation in SFM was shorter in SN12C than in 786-O (5 days vs. 7 days). Moreover, the expression levels of CD133, CD44, Nanog, and Oct3/4 in SN12C and 786-O significantly differed (P<0.05). When transplanted in nude mice, 786-O spheres were less tumorigenic than SN12C spheres. Conclusion:SN12C spheres possess the main defining characteristics of renal cancer stem cell;thus, SN12C is the more suitable cell line to be used to isolate cancer stem cells compared with 786-O.
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Objective:To investigate the effect of DKK1 on linearly patterned programmed cell necrosis (LPPCN) and vasculogenic mim-icry (VM) and the related molecular mechanism in non-small cell lung cancer (NSCLC). Methods:A total of 173 human NSCLC speci-mens were collected to detect LPPCN by H&E staining, detect VM with CD31/PAS double staining, and investigate DKK1 and related protein expression by immunohistochemistry. The clinical pathological significance of LPPCN, VM, and DKK1 and the correlation of them were analyzed. Human NSCLC H460-DKK1 cells were engrafed in nude mice to evaluate the influence of DKK1 up-regulation on VM and LPPCN in vivo. Results:Approximately, 14.45%(25/173) of NSCLC had VM and 49.71%(86/173) had LPPCN. 25.6%(22/86) of NSCLC cases in LPPCN-positive group formed VM. Both of VM and LPPCN were all correlated with poor differentiation, late TNM stage, easy recurrence and metastasis and poor prognosis in NSCLC. DKK1 expression in the VM-positive group and the LPPCN-positive group was higher than that in the VM-negative group and the LPPCN-negative group, respectively. DKK1, LPPCN, and VM were positive-ly correlated with VE-cadherin, MMP-2,β-catenin nuclear expression and Twist1. H460-DKK1 transplantation tumor model confirmed that DKK1 promotes the expression of VM and LPPCN and related proteins in NSCLC. Conclusion:The increase of theβ-catenin and Twist1 expression induced by DKK1 may promote the formation of LPPCN and VM in NSCLC.
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Objective:To determine the expression of BMP4 in hepatocellular carcinoma (HCC) and to study the role of BMP4 in inducing epithelial-mesenchymal transition (EMT) to analyze the effect of BMP4 on the migration and invasion of HCC cells. Methods: The expression of BMP4 in HCC specimens was examined by immunohistochemistry staining, and the correlations were analyzed between the expression of BMP4 and clinicopathological data. The BMP4 expression plasmid was transfected into HepG2 cells to induce exogenous overexpression of BMP4 protein. The changes of HepG2 cell morphology were detected after BMP4 transfection by using a microscope; the changes of the expression of BMP4, EMT-related protein (E-cadherin, Vimentin) in HepG2 cells were detected by Western blot after transfection of BMP4;the wound healing assay in vitro was used to detect the effects of BMP4 gene transfection on the ability of migration of HepG2 cells;the invasion assay was used to determine the role of transfection of BMP4 on the invasive potential of HepG2 cells. Results: Immunohistochemistry staining method displayed that BMP4 expression was positively associated with age, histological differentiation, stage, and poor prognosis. After BMP4 overexpression, the morphology of HepG2 cells showed significant changes from a paving stone structure with cell-cell adhesion to a fibroblastic shape, which showed typical EMT change; Western blot exhibited that the expression of E-cadherin was downregulated and the Vimentin expression was upregulated in HepG2 cells;the wound healing and invasion assay showed that the migration and invasion potentials of HepG2 cells were significantly enhanced. Conclusion: BMP4, which displayed a high expression in HCC specimens, was closely associated with clinicopathologic data, and BMP4 may promote migration and invasion of HCC cells by inducing epithelial-mesenchymal transition.
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Objective Matrix metalloproteinase-10 (MMP-10) has been shown to be highly associated with atherosclerosis.Recent studies showed that levels of MMP-10 were elevated in infarcted tissues in acute ischemic stoke.However,serum levels of MMP-10 in patients with acute ischemic stroke have never been studied previously.This study aims to investigate the serum levels of MMP-10 in patients with acute ischemic stroke,and evaluate the association of serum levels of MMP-10 with stroke subtypes based on Trial of Org 10 172 in acute stroke treatment classifications,the severity of stroke,risk factors and carotid artery plaque.Methods The circulating levels of MMP-10 were measured by enzyme linked immunosorbent assay in 194 subjects,including 109 patients who were diagnosed as acute ischemic stroke in the Department of Neurology,Shengjing Hospital,China Medical University from April to December 2012,and the 85 healthy controls.Results Patients with acute ischemic stroke had higher serum levels of MMP-10 compared with healthy controls (6.59 (6.07,7.31) μg/L vs 5.16 (3.87,5.94) μg/L,Z =8.33,P ＜ 0.01).NIHSS score had positive correlation with serum levels of MMP-10 (r =0.204,P =0.037).Classified by risk factors,we compared the MMP-10 levels of subsets,and results displayed that statistically significant difference existed between dyslipidemia subset and non-dyslipidemia subset (Z =2.07,P =0.042).MMP-10 levels had positive correlation with serum levels of LDL-cholesterol (r =0.248,P =0.040),but negative correlation with thrombin-activatable fibrinolysis inhibitor (TAFI;r =-0.208,P =0.030).The subset with unstable plaques had higher MMP-10 levels than that with stable plaque (6.62 (6.13,7.36) μg/L) vs 6.10 (6.00,6.46) μg/L,Z =2.12,P =0.034),implying the relationship of MMP-10 and atherosclerosis.Conclusions Patients with acute ischemic stroke have higher serum levels of MMP-10 compared with the healthy controls,and MMP-10 levels have positive correlation with the severity of stroke.MMP-10 is associated with the subtypes of stroke classified by risk factors,and dyslipidemia subset has higher levels of MMP-10 than that of non-dyslipidemia subset.MMP-10 has positive correlation with LDL-cholesterol,but negative correlation with TAFI.MMP-10 may be involved in the process of formation and disruption of unstable plaques,which contribute to the stenosis of arteries and onset of acute ischemic stroke.
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Objective:To investigate the clinical significance of epithelial-to-mesenchymal (EMT) in lung squamous cell carcino-ma (LSCC) and to examine the effect of EMT on the invasive and migration abilities of LSCC. Methods:Immunohistochemical stain-ing was performed to determine the expression of E-cadherin, Vimentin, and TGF-β1 in 79 LSCC patients, and the clinical significance was explored. SK-MES-1 lung squamous carcinoma cells were cultured in conditioned medium containing various concentrations of transforming growth factor-β1 (TGF-β1) for 5 and 10 days. The expression levels of E-cadherin and Vimentin were detected via West-ern blot and reverse transcription-polymerase chain reaction (RT-PCR). With different concentrations and induction times, invasion and wound healing assays were performed to evaluate the invasion and migration abilities. Results:E-cadherin expression was significantly lower, whereas Vimentin expression was significantly higher in LSCC with lymph node metastasis than in that without noda metastasis (P<0.05). In the tissues of 79 LSCC patients, TGF-β1 expression was significantly related to lymph node metastasis (P<0.05). Western blot showed that Vimentin expression was higher, whereas E-cadherin expression was lower in TGF-β1 inducing medium with 10 ng/mL SK-MES-1 cells than in the other media. RT-PCR showed similar results. Scratch test and invasion assay both showed that treat-ment of cells with cytokines markedly enhanced the migration and invasion of the cells. Conclusion:Lymph node metastasis of LSCC correlates with EMT. SK-MES-1 cells undergo EMT via TGF-β1 induction, which enhances invasion and migration.
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Objective To investigate the effects of interferon-gamma (IFN-γ) on migration,invasion and vasculogenic mimicry (VM) formation of human melanoma cell line MUM-2B. Methods MUM-2B cells were divided into three groups, control group (10%FBS in DMEM), treatment group1 (10μg/L IFN-γ) and treatment group2 (100μg/L IFN-γ). Different concentrations of IFN-γ were added in the culture medium of MUM-2B cells. Wound-healing assay and matrigel invasion assay were performed to examine the migration and invasion ability of MUM-2B cells. Three-D culture was used to observe the VM formation. The expression of vascular endothelial growth factor (VEGF) of MUM-2B cells was detected by Western blot assay. Results The result of wound-healing assay showed that the migration distance of cells was decreased in treatment groups compared with that of control group. The migration distance of cells was decreased in treatment group 2 compared with that of treatment group 1(P＜0.05). The result of matrigel invasion assay showed that the number of invaded cells was decreased in treatment groups compared with that of control group, and which was significantly decreased in treatment group2 than that of treatment group1 (P＜0.05). The result of 3-D culture showed that cells in control group can form typical VM tube-like structures, whereas cells in treatment groups cannot. Western blot assay showed that the expression of VEGF protein was significantly decreased in treatment groups compared with that of control group, and the expression of VEGF protein was significantly decreased in treatment group2 than that of treatment group 1(P＜0.05). Conclusion These data suggest that IFN-γinhibits migration and invasion of MUM-2B cells, and inhibits VM formation by down regulating VEGF expression in vitro.
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<p><b>OBJECTIVE</b>To systematically evaluate the environmental exposure information of coke oven workers, we investigated the concentration and size distribution characteristics of the particle matter (PM) in the top working area of coke oven.</p><p><b>METHODS</b>The aerodynamic particle sizer spectrometer was employed to collect the concentration and size distribution information of PM at a top working area. The PM was divided into PM ≤ 1.0 µm, 1.0 µm < PM ≤ 2.5 µm, 2.5 µm < PM ≤ 5.0 µm, 5.0 µm < PM ≤ 10.0 µm and PM>10.0 µm based on their aerodynamic diameters. The number concentration, surface area concentration, and mass concentration were analyzed between different groups. We also conducted the correlation analysis on these parameters among groups.</p><p><b>RESULTS</b>We found the number and surface area concentration of top area particulate was negatively correlated with particle size, but mass concentration curve showed bimodal type with higher point at PM = 1.0 µm and PM = 5.0 µm. The average number concentration of total particulate matter in the top working area was 661.27 number/cm³, surface area concentration was 523.92 µm²/cm³, and mass concentration was 0.12 mg/m³. The most number of particulate matter is not more than 1 µm (PM(1.0)), and its number concentration and surface area concentration accounted for 96.85% and 67.01% of the total particles respectively. In the correlation analysis, different particle size correlated with the total particulate matter differently. And the characteristic parameters of PM2.5 cannot fully reflect the total information of particles.</p><p><b>CONCLUSION</b>The main particulate matter pollutants in the top working area of coke oven is PM1.0, and it with PM(5.0) can account for a large proportion in the mass concentration of PM. It suggest that PM1.0 and PM(5.0) should be considered for occupational health surveillance on the particulate matter in the top area of coke oven.</p>
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Humans , Air Pollutants, Occupational , Coke , Occupational Exposure , Particle Size , Particulate Matter , WorkplaceABSTRACT
Objective:To investigate the correlations of Lauren classification and world health organization (WHO) classification of gastric cancer (GC) with microvascular density (MVD), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2), and p53. Methods:The clinical data of 89 patients with GC were collected. The collected specimens were categorized on the basis of Lauren classification and WHO classification. CD34/periodic acid-Schiff (PAS) double staining was performed to validate MVD. Immunohistochemistry was conducted to investigate the expression levels of MMP-2, MMP-9, VEGF, VEGFR1, VEGFR2, and p53. Results:MVD was not correlated with Lauren classification or WHO classification (P>0.05). Lauren typing was associated with the expression levels of MMP-9, VEGFR1, and p53 (P0.05). Cox proportional hazards model revealed that Lauren classification and WHO classification were the prognostic factors of overall survival (P<0.05). Conclusion:This research on tumor related factors, angiogenesis, and different classifications of GC may provide new methods to treat this disease.
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Objective:This study aims to investigate the potential of colon cancer cells to differentiate into vascular endothelial cells in endothelial-induced specific environment. Methods:Three colon cancer cells with different differentiated level HCT116 (poor-ly differentiated), SW480 (moderately differentiated), HT29 (well differentiated) were cultured in the conditioned medium containing the endothelial-inducing factors for 15 days respectively. The expression of vascular endothelial indicators Platelet endothelial cell adhe-sion molecule-1、Endothelial cell adhesion molecule CD34 was detected via western blot. Immunofluorescence staining was performed to examine CD31 and CD34 expression level in HCT116 after cultured in endothelial-inducing medium and ordinary medium for 15 days respectively, and the three-dimensional (3D) culture was used to detect the abililty of in vitro tube-like structure formation. Re-sults:Western blot showed that CD31 and CD34 expression level were negatively correlated with degree of differentiation in colon can-cer cells. CD31 and CD34 expression in endothelial-inducing medium HCT116 cells (poorly differentiated) were higher then in the nor-mal medium, while the CD31 and CD34 expression in SW480 cells (moderately differentiated) and HT29 cells (well differentiated) in the two cultural mediums were not notably changed. Immunofluorescence staining illustrated that CD31 and CD34 expression in HCT116 cells cultured in endothelial-inducing medium increased compared with those cultured in ordinary medium. In vitro three-di-mensional culture demonstrated that ability of tube-like structure formation was notably enhanced after endothelial-inducing cultured. Conclusion:Endothelial-inducing medium could promote colon cancer cells with strong stemness differentiate toward vascular endo-thelial cells.
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<p><b>OBJECTIVE</b>To evaluate the effectiveness of resin-based sealants and glass ionomer sealants on caries prevention in the first permanent molars.</p><p><b>METHODS</b>A randomized controlled, examiner-blinded, 2-year clinical trial with parallel groups was conducted. A total of 419 children (664 first molars) who had at least one sound first permanent molar with deep fissures or fissures with signs of non-cavitated lesions were recruited. They were randomly allocated into three groups.Helioseal F, a light-cured resin-based sealants was used in group R (136 children with 219 first molars). FUJI VII, a light-cured glass ionomer sealants was used in group G (130 children with 218 first molars) and group N (153 children with 227 first molars) was blank control.Follow-up examinations were conducted at 6, 12, and 24 month after sealing to examine the sealants retention and caries incidence. Chi-square test was used to compare the differences of caries incidence among the 3 groups.Generalized estimating equations (GEE) was used to analyze the effective factors of caries incidence.</p><p><b>RESULTS</b>During the 2-year follow-up period, 3.0% (6/197) of the teeth in group R, 6.7% (13/193) of the teeth in group G, and 14.7% (29/197) of the teeth in group N were diagnosed as having developed a cavitated lesion. The difference in the incidence of caries among the three groups was significant(P < 0.05). No significant difference in caries incidence was found between group R and group G (P > 0.05). The tooth-level preventive fractions were 79.6% for resin-based sealants and 54.4% for glass ionomer sealants, respectively.</p><p><b>CONCLUSIONS</b>Both resin-based sealants and glass ionomer sealants can effectively prevent the incidence of caries in the first permanent molars. The two sealants have similar caries prevention effect.</p>
Subject(s)
Child , Humans , Acrylic Resins , Composite Resins , Dental Caries , Glass Ionomer Cements , Molar , Pit and Fissure Sealants , Silicon Dioxide , Single-Blind MethodABSTRACT
The aim of this research was to explore the most optimal method of DNA extraction from formalin-fixed, paraffin-embedded (FFPE) tissues, and to improve the amplification of long fragments with the method. Three methods, one step method, phenol-chloroform extraction method, and genomic DNA purification kit method, were employed to extract DNA from twenty normal thyroid tissues which were fixed with formalin and embedded with paraffin. The highest proportionality of OD260/OD280 in the examples was obtained by phenol-chloroform extraction method, 1.703 +/- 0.086, compared to the results of the other two methods. As for the long DNA segments amplification, the achievement ratio of one step method, phenol-chloroform extraction method and genomic DNA purification kit method were 0%, 5% and 10%, respectively, by traditional PCR method, but 0%, 95% and 85% respectively by the nest PCR. We have found that the best process of extracting DNA from FFPE is digesting by proteinase K and purifying by phenol-chloroform, and it is effective to amplify long DNA segments from FFPE by nest PCR.