Study on the mechanism of fatty acid binding protein 5 binding Vimentin protein in the Hepatocellular Carcinoma cells / 实用医学杂志

Objective The aim of this study was to screen and verify the proteins interacting with Vimentin,investigate the regulatory relationship between FABP5 and candidate proteins,and further explore the mechanism of FABP5 in hepatocellular carcinoma.Methods Immunoprecipitation combined with tandem mass spectrometry(IP-MS)was used to screen the proteins that bind to FABP5.The binding relationship between FABP5 and candi-date interacting proteins was verified from the exogenous and endogenous levels by Co-immune precipitation assay(Co-IP).RT-qPCR,Western blot and immunofluorescence were used to observe the effect of knockdown FABP5 on the transcription and translation of Vimentin in HCC cells.The effect of overexpressing FABP5 on the cytoskeleton of HCC cell was observed by phalloidin staining.Results 336 potential target proteins that bind to FABP5 were identi-fied through IP-MS.Based on literature,five candidate proteins related to tumors were selected,namely PRDX1,PRSS3,PKM,HSP90AA1,and Vimentin.The binding relationship between FABP5 and Vimentin protein was con-firmed through both exogenous and endogenous Co-IP.Knockdown FABP5 has no significant effect on the expression of Vimentin mRNA,but it can inhibit the expression of Vimentin protein,and overexpression of FABP5 can affect the cytoskeleton of HCC cell.Conclusions FABP5 promotes the migration and invasion of HCC cells by the regula-tion of Vimentin and the influence of cytoskeletal remodeling,and thus it is expected to be a potential target for anti-HCC and provide new ideas for the treatment of HCC.

Role and mechanism of vimentin in neonatal meningitis induced by group B streptococcus / 中国生物制品学杂志

@#Objective To evaluate the role of vimentin(Vim)in neonatal meningitis induced by group B streptococcus(GBS)and explore its molecular mechanism,so as to provide experimental basis for the prevention and treatment of GBS neonatal meningitis. Methods Human brain microvascular endothelial cells(HBMECs)were used as the cell model,the Vim knockdown strain was constructed by transfection of Vim small interfering RNA(siRNA)sequence into HBMECs,and NC siRNA group transfected with negative control sequence was set at the same time. The inhibitory effect of Vim was detected by Western blot and immunofluorescence assay. After 48 h of transfection,the cells were infected with GBS at a MOI of 100,and adhesion and invasion tests were performed 2 h and 3 h after infection,respectively. After 3 h of infection,the effect of Vim inhibition on NF-κB p65 nuclear translocation in HBMECs was measured by Western blot and immunofluorescence assay,and the effect of Vim inhibition on ERK pathway activation related proteins and inflammatory factors in HBMECs was detected by Western blot. Results Compared with the control group,the Vim protein level and fluorescence intensity in HBMECs of Vim siRNA group significantly decreased(t = 3. 242 and 71. 51,P < 0. 05 and < 0. 001,respectively). Compared with NC siRNA + GBS group,the adhesion rate and invasion rate of Vim siRNA + GBS group decreased significantly(t = 9. 949 and 30. 050,respectively,each P < 0. 001);the phosphorylated p65 level decreased significantly(t = 2. 824,P < 0. 05),the nuclear translocation of p65 protein was inhibited,and the intensity of green fluorescence in nucleus decreased significantly;the levels of TNF-α,IL-6 and phosphorylated ERK1/2 proteins were significantly inhibited(t = 4. 000,6. 367 and 3. 872,respectively,each P < 0. 05). Conclusion Inhibition of Vim can reduce the adhesion and invasion of GBS to HBMECs,inhibit the activation of NF-κB and ERK signaling pathway and the production of downstream inflammatory factors,thus reducing the damage of GBS to HBMECs.

Imunoexpressão de clic4, α-sma, e-caderina e vimentina em lesões odontogênicas epitelias benígnas / Immunoexpression of clic4, α-sma, e-cadherin and vimentin in benign epithelial odontogenic lesions

As lesões odontogênicas epiteliais benignas constituem um grupo heterogêneo de lesões. A proteína CLIC4 atua na regulação dos processos de parada de crescimento e apoptose, participando também do processo de transdiferenciação dos fibroblastos em miofibroblastos que passam a expressar α-SMA. Além disso, a expressão de CLIC4 pode interferir no processo de transição epitélio-mesenquima (TEM) em neoplasias. Este trabalho avaliou a imunoexpressão de CLIC4, α-SMA, E-caderina e Vimentina em ameloblastomas (AM) (n = 16), ceratocistos odontogênicos (n = 20) e tumores odontogênicos adenomatóides (TOA) (n = 8). A análise da expressão imunoistoquímica das proteínas CLIC4, E-caderina e vimentina no componente epitelial das lesões e de CLIC4 e α-SMA no tecido conjuntivo foi realizada de forma semi-quantitativa por um avaliador previamente calibrado. A expressão no componente epitelial de CLIC4 foi analisada separadamente no núcleo e no citoplasma, bem como a marcação de E-caderina que foi avaliada na membrana e no citoplasma. As comparações dos percentuais de imunorreatividade em relação aos grupos estudados foram realizadas por meio dos testes não paramétricos de Kruskal-Wallis e Mann-Whitney. Possíveis correlações entre a expressão de CLIC4, α-SMA, E-caderina e Vimentina foram avaliadas por meio do teste de correlação de Spearman. O nível de significância foi estabelecido em 5% (p < 0,05). Foram observados diferentes padrões de marcação entre os grupos analisados, observando-se que a imunoexpressão exclusivamente citoplasmática da CLIC4 no componente epitelial dos AM (p < 0,001) e TOA (p < 0,001) foi significativamente superior a dos CO, não demonstrarando significância estatística entre os AM e TOA. A imunoexpressão (nuclear e citoplasmática) da CLIC4 no revestimento epitelial CO foi significativamente superior à encontrada no componente epitelial dos AM (p < 0,001) e dos TOA (p < 0,001). A imunoexpressão estromal de CLIC4 foi significativamente superior nos AM (p = 0,009) e CO (p = 0,004) quando comparados aos TOA. A imunoexpressao de α-SMA significativamente maior em AM (p = 0,016) e CO (p = 0,034) quando comparados aos TOA. Para a imunoexpressão membranar da E-caderina em CO foi significativamente superior em comparação à encontrada nos AM (p = 0,009) e nos TOA (p = 0,024). Foi observada maior imunoexpressão de E-caderina (membranar e citoplasmática) nos COs, quando comparados aos AM (p < 0,001) e aos TOAs (p < 0,001). A expressão de Ecaderina citoplasmática foi significativamente maior nos AM e TOA (p < 0,001) quando comparados aos CO. Observou-se diferença estatisticamente significativa na imunoexpressão de vimentina entre os casos de AM e os casos de TOA (p = 0,038) e CO (p < 0,001), bem como entre o TOA e CO (p < 0,001). As correlações testadas entre os escores das proteínas estudadas evidenciou que no grupo dos AM foi possível evidenciar moderada correlação positiva e estatisticamente significativa (r = 0,527; p = 0,036) entre a expressão citoplasmática da CLIC4 e a expressão citoplasmática da E-caderina. Também foi verificada fraca correlação negativa e estatisticamente significativa (r = -0,499; p = 0,049) entre a expressão núcleo-citoplasmática da CLIC4 e a expressão citoplasmática da E-caderina nos AM. Além disso, uma moderada correlação positiva e estatisticamente significativa entre a expressão estromal da CLIC4 e a expressão da α-SMA nos AM (r = 0,648; p = 0,007) e nos CO (r = 0,541; p = 0,014). Foi observada forte correlação negativa e estatisticamente significativa (r = -0,813; p < 0,001) entre a expressão da E-caderina e a expressão da vimentina nos AM. Os resultados deste estudo sugerem um potencial envolvimento de CLIC4 no processo de transdiferenciação de miofibroblastos, e que a presença destas células é mais frequentemente associada a lesões de comportamento biológico mais agressivo como os AM e CO, além de uma possível atuação desta proteína na regulação do ciclo celular e na TEM nas lesões estudadas (AU).

Benign epithelial odontogenic lesions constitute a heterogeneous group of lesions. the CLIC4 protein acts in the regulation of growth arrest and apoptosis processes, also participating in the process of transdifferentiation of fibroblasts Into myofibroblasts that begin to express α-SMA. Furthermore, CLIC4 expression can interfere with the epithelialmesenchymal transition (EMT) process in neoplasms. This work evaluated the immunoexpression of CLIC4, α-SMA, e-cadherin and vimentin in ameloblastomas (AM) (n = 16), odontogenic keratocysts (OK) (n = 20) and adenomatoid odontogenic tumors (AOT) (n = 8). The analysis of the immunohistochemical expression of the proteins CLIC4, ecadherin and vimentin in the epithelial component of the lesions and of CLIC4 and α-SMA in the connective tissue was carried out in a semi-quantitative way by a previously calibrated evaluator. Expression in the epithelial component of CLIC4 was analyzed separately in the nucleus and cytoplasm, as well as e-cadherin labeling, which was evaluated in the membrane and cytoplasm. Comparisons of the percentages of immunoreactivity in relation to the studied groups were carried out using the nonparametric kruskal-wallis and mann-whitney tests. Possible correlations between the expression of CLIC4, α-SMA, e-cadherin and vimentin were evaluated using the spearman correlation test. The significance level was set at 5% (p < 0.05). Different staining patterns were observed between the groups analyzed, observing that the exclusively cytoplasmic immunoexpression of CLIC4 in the epithelial component of AM (p < 0.001) and AOT (p < 0.001) was significantly higher than that of OK, not demonstrating statistical significance between the AM and AOT. The immunoexpression (nuclear and cytoplasmic) of CLIC4 in the co epithelial lining was significantly higher than that found in the epithelial component of AM (p < 0.001) and AOT (p < 0.001). Stromal CLIC4 immunoexpression was significantly higher in AM (p = 0.009) and OK (p = 0.004) when compared to AOT. The immunoexpression of α-SMA is significantly higher in AM (p = 0.016) and OK (p = 0.034) when compared to AOT. For e-cadherin membrane immunoexpression in co was significantly higher compared to that found in AM (p = 0.009) and AOT (p = 0.024). Greater immunoexpression of e-cadherin (membrane and cytoplasmic) was observed in OK, when compared to AM (p < 0.001) and AOT (p < 0.001). Cytoplasmic ecadherin expression was significantly higher in AM and AOT (p < 0.001) when compared to OK. A statistically significant difference in vimentin immunoexpression was observed between cases of AM and cases of AOT (p = 0.038) and OK (p < 0.001), as well as between AOT and OK (p < 0.001). The correlations tested between the scores of the proteins studied showed that in the am group it was possible to demonstrate a moderate positive and statistically significant correlation (r = 0.527; p = 0.036) between the cytoplasmic expression of clic4 and the cytoplasmic expression of e-cadherin. A weak and statistically significant negative correlation (r = -0.499; p = 0.049) was also found between the nucleus-cytoplasmic expression of clic4 and the cytoplasmic expression of e- cadherin in AM. Furthermore, a moderate positive and statistically significant correlation between the stromal expression of CLIC4 and the expression of α-SMA in AM (r = 0.648; p = 0.007) and OK (r = 0.541; p = 0.014). Additionally, a strong negative and statistically significant correlation (r = -0.813; p < 0.001) was observed between the expression of ecadherin and the expression of vimentin in AM. The results of this study suggest a potential involvement of CLIC4 in the myofibroblast transdifferentiation process, and that the presence of these cells is more frequently associated with lesions with more aggressive biological behavior such as AM and OK, in addition to a possible role of this protein in the regulation of cell cycle and EMT in the lesions studied (AU).

Correlation Between Vimentin and Hepatocellular Carcinoma and Intervention Effect of Jianpi Yiqi Prescription / 中国实验方剂学杂志

ObjectiveTo study the correlation between Vimentin and hepatocellular carcinoma （HCC） and the mechanism of Jianpi Yiqi prescription against HCC through Vimentin. MethodCorrelation between Vimentin and HCC was analyzed based on the cancer genome atlas（TCGA）， clinical proteomic tumor analysis consortium（CPTAC）， STRING， and Cytoscape. SD rats were randomized into normal group （normal saline， ig， once/day， 4 weeks）， model group （normal saline， ig， once/day， 4 weeks）， low-dose， medium-dose， and high-dose （5.25， 10.5， 21 g·kg-1， ig， once/day， 4 weeks） JianpiYiqi prescription groups， signal transducer and activator of transcription 3 （STAT3） inhibition group （C188-9， 4.5 mg·kg-1， ip， once/day， 4 weeks）， and glycoprotein 130 （gp130） inhibition group （SC144， 4.5 mg·kg-1， ip， once/day， 4 weeks）， 10 rats in each group. Diethylnitrosamine （DEN， 70 mg·kg-1 body weight， ip） was injected in rats except the normal group to induce HCC. After the modeling， administration began. After last administration， Real-time polymerase chain reaction（Real-time PCR） was performed to determine Vimentin mRNA level in rat liver tissue. Caspase-3 activity in liver tissue was detected by colorimetry， and expression of Rho kinase （ROCK）1， ROCK2， aurora kinase B （AURKB）， Zinc-finger protein 148 （ZNF148）/zinc-binding protein-89 （ZBP-89）， STAT3， p-STAT3， total Vimentin， and phosphorylated （p）-Vimentin in liver tissue and Vimentin in liver tissue nucleus detected by Western blot. Serum Vimentin concentration was measured by enzyme-linked immunosorbent assay （ELISA）. ResultVimentin mRNA level was high in tissues from HCC patients with different cancer stages （stage Ⅰ-Ⅳ）， different pathological grades （G1-G3）， no regional lymph node metastasis （N0）， and different subtypes （P<0.01）. Vimentin mRNA expression was higher in tissues from patients with lymph node metastasis than in patients without lymph node metastasis and normal samples. Vimentin protein level was decreased in HCC tissues （P<0.01）. Vimentin gene has 4 mutations which can induce change in the primary structure of Vimentin protein and patients with Vimentin gene mutation had short disease free survival time （P<0.01）. The mRNA expression of Vimentin was negatively associated with HCC cell purity （P<0.01） but was positively associated with the infiltration levels of cancer-associated fibroblasts， M2 macrophages， myeloid dendritic cell and other immune cells in tumor microenvironment （P<0.01）. Association analysis results showed that the expression of Vimentin was correlated with the STAT3 expression in HCC tissues （P<0.01）. As for the animal experiment， Vimentin mRNA level and protein levels of total Vimentin and p-Vimentin in liver tssue， Vimentin protein level in liver tissue nucleus， Vimentin in rat serum， ROCK2， AURKB， STAT3 and p-STAT3 in liver tissues were up-regulated （P<0.01） and protein level of negative regulator ZBP-89 was reduced in the model group （P<0.01） compared with those in the normal group. Activity of Caspase-3 in liver tissue increased and the ROCK1 protein level was increased in the model group compared with those in the normal group. STAT3 inhibitor， gp130 inhibitor， and medium-dose and high-dose Jianpi Yiqi prescription all can reduce the secretory Vimentin protein in serum， protein levels of total Vimentin and p-Vimentin in liver tissues， and Vimentin in liver tissue nucleus， and the protein levels of STAT3/Vimentin signaling pathway-related molecules， such as STAT3， p-STAT3， ROCK2， and AURKB and up-regulate the protein level of negative regulator ZBP-89 and activity of Caspase-3 （P<0.05， P<0.01）. Effect of medium-dose or high-dose Jianpi Yiqi prescription on Vimentin mRNA expression， STAT3 protein expression， ZBP-89 protein expression， ROCK2 protein expression， AURKB protein expression and Caspase-3 activity was not significantly different from that of STAT3 inhibitor. ConclusionVimentin， an important inflammatory molecule， is closely related to the occurrence and development of HCC and its expression， subcellular location and function may be affected by cancer-associated fibroblasts， M2 macrophages， myeloid dendritic cell， and IL-6/STAT3 signaling pathway， particularly by STAT3 molecule. Jianpi Yiqi prescription may exert therapeutic effect on HCC via regulating Vimentin through the STAT3/Vimentin signaling pathway.

Berberine Promotes Mouse Breast Cancer Metastasis to Lung via Inducing Epithelial–mesenchymal Transition / 肿瘤防治研究

Objective To study the effect and mechanism of berberine (BBR) on the lung metastasis of mouse breast cancer via epithelial-mesenchymal transition (EMT). Methods CCK-8 and Transwell migration assays were utilized to investigate the proliferation and migration properties of breast cancer 4T1 cells after BBR treatment.Mouse 4T1-Luc cells were injected into mice under the fourth mammary fat pad, and the mice were then randomly divided into the control and BBR groups.The mice in the BBR group received daily intraperitoneal injections of BBR working solution and those in the control group were continuously intraperitoneally injected with the same volume of the solvent used to dissolve BBR powder.Tumor metastasis in the lungs of living mice was detected by using an in vivo imaging system.After 42 days of administration, lung metastasis was measured via microscopy and HE staining.Western blot analysis was used to examine the effects of BBR on the expression of EMT-related proteins (Vimentin and Snail) as well as the activation of the Akt and ERK signaling pathways. Results BBR significantly promoted 4T1 cell migration (P < 0.05).In vivo experiments showed that the number of lung metastases in the BBR group had significantly increased compared with that in control group (P < 0.05) as observed under microcopy and histological staining.Compared with the control group, BBR upregulated the expression levels of Vimentin and Snail as well as the phosphorylated levels of p-Akt and p-ERK (P < 0.05). Conclusion BBR may promote EMT and lung metastasis of breast cancer 4T1 cells by activating the expression of proteins in the p-Akt and p-ERK pathways.

Study on Jianpi Bushen Jiedu Decoction for inhibiting epithelial-mesenchymal transformation of hepatocellular carcinoma through JAK2/STAT3 pathway / 国际中医中药杂志

Objective:To study the effects of Jianpi Bushen Jiedu Decoction on the epithelial-mesenchymal transformation of nude mice with HCCLM3 subcutaneous transplanted tumor by regulating JAK2/STAT3 pathway.Methods:HCCLM3 subcutaneous transplanted tumor model was established in mice. After the successful modeling, 24 nude mice were divided into blank group, TCM group and combined group according to random number table method, with 8 mice in each group. Mice in the TCM group were given 0.68 mg/ml alcohol extract of Jianpi Bushen Jiedu Decoction for gavage, and the combined group were given sorafenib suspension plus alcohol extract of Jianpi Bushen Jiedu Decoction 3.5 mg/ml for gavage, once a day, for consecutive 4 weeks. The effects of Jianpi Bushen Jiedu Decoction on tumor volume, tumor weight of HCCLM3 subcutaneous transplanted tumor and mice body weight were observed; Western blot was used to detect the expressions of E-cadherin, N-cadherin, Vimentin and JAK2/STAT3 pathway-related proteins in subcutaneous transplanted tumor tissues of hepatocellular carcinoma of mice in each group.Results:Compared with the control group, the average tumor weight of subcutaneous transplanted tumor decreased significantly in the TCM group and the combined group ( P<0.05), and the expressions of JAK2, STAT3, p-JAK2, p-STAT3, N-cadherin, and Vimentin decreased significantly in subcutaneous transplanted tumor tissue ( P<0.05), while E-cadherin increased ( P<0.05). Conclusion:Jianpi Bushen Jiedu Decoction can inhibit the growth of subcutaneous transplanted tumor of hepatocellular carcinoma in mice. The mechanism may be related to inhibiting the activation of JAK2/STAT3 pathway, thereby inhibiting the epithelial-mesenchymal transformation of hepatocellular carcinoma.

The expression and significance of Piezo1 in chronic rhinosinusitis with nasal polyps / 临床耳鼻咽喉头颈外科杂志

Objective:To explore the expression and importance of Piezo1, E-cadherin, and Vimentin in nasal polyps patients. Methods:Thirty-five patients undergoing endoscopic sinus surgery under general anesthesia were streamed into 20 cases of nasal polyps（NP group） and 15 cases of simple septoplasty without any sinus disease（Control group）. Immunofluorescence staining and Western Blot were applied to detect the protein level of Piezo1, E-cadherin, and Vimentin in NP tissues and nasal polyp-derived primary human nasal epithelial cells（pHNECs）. Also, BEAS-2B cell lines were treated with human TGF-β1 protein to establish epithelial mesenchymal transition（EMT） model in vitro and quantitative real-time polymerase chain reaction were used to calculate Piezo1 and above biomarkers in the model. Results:Compared with control group, Piezo1 and Vimentin showed higher level while E-cadherin was lower in NP tissues and pHNECs.In EMT model in vitro, Piezo1 and Vimentin were demonstrated higher expression with decreased level of E-cadherin. Conclusion:The tendency of Piezo1 is consistent with the mesenchymal-related biomarker Vimentin, going against with epithelial-related biomarker E-cadherin, implying its involvement with EMT process in nasal polyps.

Estudo da imunoexpressão de proteínas envolvidas na transição epitélio-mesênquima em tumores de glândula salivar / Study of the immunoexpression of proteins involved in the epithelial-mesenchymal transition in salivary gland tumors

Os tumores de glândula salivar (TGS) apresentam notável complexidade clínica e biológica, razão para a qual muitos estudos investigam os eventos envolvidos na sua progressão. Uma das dinâmicas envolvidas na invasão tumoral de diversos tipos de carcinomas é a transição epitélio-mesênquima (TEM). Neste processo, as células epiteliais sofrem transição para um estado mesenquimal móvel, favorecendo a invasão e metástase. Sendo assim, esta pesquisa analisou a expressão imuno-histoquímica de E-caderina, Twist1, Snail1, α-SMA, metaloproteinases de matriz 9 (MMP-9) e Vimentina (VM) em 90 casos de TGS, correlacionando-os entre si e com parâmetros clinicopatológicos. Foram selecionados 20 casos de Adenoma pleomórfico (AP), 20 casos de Carcinoma mucoepidermoide (CME), 20 casos de Carcinoma adenoide cístico (CAC), 10 casos de Adenocarcinoma polimorfo (ACP), 10 casos de Carcinoma epitelial-mioepitelial (CEME) e 10 casos de Carcinoma ex-adenoma pleomórfico (CexAP). A análise de E-caderina, Twist1, Snail1 foi realizada em parênquima tumoral sendo observado o percentual de células positivas (PP), com escores variando de 0 a 4, e a intensidade de expressão (IE), cujos escores variaram de 0 a 3. A avaliação de MMP-9 foi realizada em parênquima e estroma tumoral, também avaliando-se a PP e a IE, ambos baseados em escores que variaram de 0 a 3. A marcação para α-SMA e VM foi analisada em região de estroma tumoral. Células positivas para α-SMA foram contabilizadas em 10 campos, obtendo-se, então a média. A VM foi avaliada de forma qualitativa, utilizando-se 4 escores de acordo com a IE e se a marcação é difusa ou focal. Os dados obtidos foram analisados no software Statistical Package for Social Science, GraphPad Prism e STATA. O nível de significância de 5% foi adotado para os testes estatísticos. Foi verificada menor imunomarcação de E-caderina nos APs em relação às neoplasias malignas de glândula salivar (NMGS). Observou-se baixa imunoexpressão de Twist1 e Snail1 em APs. Em relação a expressão nuclear do Twist1, constatou-se maior expressão nas neoplasias malignas quando comparadas aos APs. Ainda, Twist1 em núcleo foi correlacionado à expressão citoplasmática de E-caderina nas NMGS. No que concerne aos parâmetros clinicopatológicos, esta proteína se relacionou estatisticamente com maiores chances de óbito. Foi evidenciada baixa imunoexpressão de Snail1 entre as NMGS. No entanto, na análise dos CACs, foi verificada maior expressão nuclear na variante sólida em relação às demais. A expressão de MMP-9 em parênquima demonstrou correlação positiva com Twist1 citoplasmático e Snail1nuclear nas NMGS. A MMP-9 também apresentou correlação positiva na comparação da sua imunoexpressão em região de parênquima e de estroma. A VM se apresentou como um biomarcador a ser considerado na avaliação clínica dos pacientes, já que esta apresentou relação significativa com tamanho do tumor (T3-T4) e maior frequência de óbito. Ademais, a alta expressão desta proteína se apresentou como um fator preditivo independente para piores taxas de sobrevida global (SG). A avaliação dos demais fatores clinicopatológicos apresentou estágios clínicos avançados como indicador de valor prognóstico independente para menores taxas de SG, enquanto que para a sobrevida livre da doença, estes foram a localização em glândula salivar menor e presença de metástase à distância. Os resultados deste estudo sugerem que o processo de TEM pode estar relacionado ao estágio de diferenciação celular em APs e à progressão tumoral nas NMGS. Ressalta-se, também, maior participação de Twist1 e MMP-9 no cenário da TEM em tumores malignos de glândula salivar, além da possibilidade de utilização da VM como indicador de valor prognóstico (AU).

Salivary gland tumors (SGTs) present remarkable clinical and biological complexity; therefore, many studies investigate the events involved in their progression. One of the dynamics involved in the tumor invasion of different types of carcinomas is the epithelial-mesenchymal transition (EMT). In this process, epithelial cells undergo a transition to a mobile mesenchymal state, favoring invasion and metastasis. Therefore, this research analyzed the immunohistochemical expression of E-cadherin, Twist1, Snail1, α-SMA, vimentin (VM) and matrix metalloproteinase 9 (MMP-9) in 90 SGTs cases; correlations among the biomarkers, as well as between the biomarkers and clinicopathological parameters were made. We selected 20 cases of pleomorphic adenoma (PA), 20 cases of mucoepidermoid carcinoma (MEC), 20 cases of adenoid cystic carcinoma (ACC), 10 cases of polymorphous adenocarcinoma (PAC), 10 cases of epithelial-myoepithelial carcinoma (EMC) and 10 cases of carcinoma ex-pleomorphic adenoma (CXPA). E-cadherin, Twist1, and Snail1 were analyzed in tumor parenchyma, observing the percentage of positive cells (PP) using scores ranging from 0 to 4, and the expression intensity (EI), whose scores were ranged from 0 to 3. The evaluation of MMP-9 was performed in tumor parenchyma and stroma, also evaluating PP and IE, both based on scores that ranged from 0 to 3. The labeling for α-SMA and VM was analyzed in stromal cells. Positive cells for α-SMA were counted in 10 fields and the mean was calculated. VM was evaluated qualitatively, using 4 scores according to EI and whether the labeling was diffuse or focal. Obtained data were analyzed using Statistical Package for Social Science, GraphPad Prism, and STATA software. The significance level of 5% was adopted for the statistical tests. Patients were mostly female, with a mean age of 49.8 years; the major salivary glands were the most affected anatomical site, mainly the parotid gland. A lower E-cadherin immunostaining was verified in PAs in comparison to malignant neoplasms of salivary glands (MNSGs). Low immunoexpression of Twist1 and Snail1 was observed in PAs. Regarding the nuclear expression of Twist1, it was found greater expression in malignant neoplasms than in PAs. Furthermore, Twist1 in the nucleus was correlated with cytoplasmic expression of E-cadherin in MNSGs. Regarding clinicopathological parameters, this protein was statistically related to higher chances of death. Low immunoexpression of Snail1 was evidenced among the MNSGs. However, in the analysis of CACs, greater nuclear expression was observed in the solid variant compared to the others. Expression of MMP-9 in parenchyma showed a positive correlation with cytoplasmic Twist1 and Snail1nuclear in MNSGs. MMP-9 also showed a positive correlation when comparing its immunoexpression in the parenchyma and the stroma. VM was presented as a biomarker to be considered in the clinical evaluation of patients since it showed a significant correlation between greater tumor size and a higher frequency of death. Furthermore, the high expression of this protein appeared as an independent predictive factor for worse overall survival (OS) rates. The evaluation of the rest of the clinicopathological factors showed advanced clinical stages as an indicator of independent prognostic value for lower rates of OS. For disease-free survival, these indicators were the location in the minor salivary gland and the presence of distant metastasis. Our results suggest that the EMT may be related to myoepithelial differentiation in PAs and tumor progression in MNSGs. Also, Twist1 and MMP-9 appear to play a greater role in the scenario of EMT in MNSGs; finally, VM might be used as a prognostic value indicator (AU).

Low-dose cisplatin exposure and SNAIL, Vimentin, E-cadherin expression in HEPG2 cell line / Exposición a dosis bajas de cisplatino y expresión de SNAIL, Vimentina, E-cadherina en línea celular HEPG2

Cisplatin, the first platinum compound approved for cancer treatment, is widely used in the treatment of various cancers including hepatocellular carcinoma (HCC). HCC incidence rates rise globally. Epithelial mesenchymal transition (EMT) is implicated in cancer invasion and metastasis, which are associated with increased mortality. Cisplatin dose might influence cancer invasion and metastatic behavior of the cells. The aim of the study was to investigate the effect of low-dose cisplatin treatment on EMT- related changes in HepG2 cells. Following treatment with 4 µM cisplatin, HepG2 cells were evaluated morphologically. Gene expression of E-cadherin, Vimentin, Snail1 was assessed by quantitative PCR. Immunofluorescence analyses of NA-K ATPase were performed. Although the low-dose cisplatin treated cells exhibited a more stretched morphology, no statistical difference was detected in gene expression of E-cadherin, Vimentin, Snail1 and immunofluorescence of NA-K ATPase. Findings on low-dose cisplatin effects in HepG2 might contribute to the knowledge of antineoplastic inefficacy by further understanding the molecular mechanisms of drug action.

El cisplatino, el primer compuesto de platino aprobado para el tratamiento del cáncer, es ampliamente utilizado en el tratamiento de varios tipos de cáncer, incluido el carcinoma hepatocelular (CHC). Las tasas de incidencia de CHC aumentan a nivel mundial. La transición mesenquimal epitelial (EMT) está implicada en la invasión del cáncer y la metástasis, que se asocian con un aumento de la mortalidad. La dosis de cisplatino podría influir en la invasión del cáncer y el comportamiento metastásico de las células. El objetivo del estudio fue investigar el efecto del tratamiento con dosis bajas de cisplatino en los cambios relacionados con la EMT en las células HepG2. Tras el tratamiento con cisplatino de 4 µM, se evaluaron morfológicamente las células HepG2. La expresión génica de E-cadherina, vimentina, caracol1 se evaluó mediante PCR cuantitativa. Se realizaron análisis de inmunofluorescencia de NA-K ATPasa . Aunque las células tratadas con cisplatino en dosis bajas exhibieron una morfología más estirada, no se detectaron diferencias estadísticas en la expresión génica de E-cadherina, vimentina, Snail1 e inmunofluorescencia de NA-K ATPasa. Los hallazgos sobre los efectos del cisplatino en dosis bajas en HepG2 podrían contribuir al conocimiento de la ineficacia antineoplásica al comprender mejor los mecanismos moleculares de la acción del fármaco.

Mitochondrial energy metabolism in baby hamster kidney (BHK-21/C13) cells treated with karnozin extra® / Metabolismo energético mitocondrial en células de riñón de hámster bebé (BHK-21 / C13) tratadas con karnozin extra®

SUMMARY: Carnosine is known as a natural dipeptide, which inhibits the proliferation of tumor cells throughout its action on mitochondrial respiration and cell glycolysis. However, not much is known about its effects on the metabolism of healthy cells. We explored the effects of Karnozin EXTRA® capsule with different concentrations of L-carnosine, on the cell viability and the expressions of intermediate filament vimentin (VIM) and superoxide dismutase (SOD2) in normal fibroblasts BHK-21/C13. Furthermore, we investigated its action on the energy production of these cells. Cell viability was quantified by the MTT assay. The Clark oxygen electrode (Oxygraph, Hansatech Instruments, England) was used to measure the "intact cell respiration rate", state 3 of ADP-stimulated oxidation, maximum oxidation capacity and the activities of complexes I, II and IV. Results showed that Karnozin EXTRA® capsule in concentrations of 2 and 5 mM of L-carnosine did not induce toxic effects and morphological changes in treated cells. Our data revealed a dose-dependent immunofluorescent signal amplification of VIM and SOD2 in the BHK-21/C13 cell line. This supplement substantially increased the recorded mitochondrial respiration rates in the examined cell line. Due to the stimulation of mitochondrial energy production in normal fibroblasts, our results suggested that Karnozin EXTRA® is a potentially protective dietary supplement in the prevention of diseases with altered mitochondrial function.

RESUMEN: La carnosina se conoce como dipéptido natural, que inhibe la proliferación de células tumorales a través de su acción sobre la respiración mitocondrial y la glucólisis celular. Sin embargo, no se sabe mucho de sus efectos sobre el metabolismo de las células sanas. Exploramos los efectos de la cápsula Karnozin EXTRA® con diferentes concentraciones de L-carnosina, sobre la viabilidad celular y las expresiones de vimentina de filamento intermedio (VIM) y superóxido dismutasa (SOD2) en fibroblastos normales BHK-21 / C13. Además, estudiamos su acción sobre la producción de energía de estas células. La viabilidad celular se cuantificó mediante el ensayo MTT. Se utilizó el electrodo de oxígeno Clark (Oxygraph, Hansatech Instruments, Inglaterra) para medir la "tasa de respiración de células intactas", el estado 3 de oxidación estimulada por ADP, la capacidad máxima de oxidación y las actividades de los complejos I, II y IV. Los resultados mostraron que la cápsula de Karnozin EXTRA® en concentraciones de 2 y 5 mM de L- carnosina no indujo efectos tóxicos ni cambios morfológicos en las células tratadas. Nuestros datos revelaron una amplificación de señal inmunofluorescente dependiente de la dosis de VIM y SOD2 en la línea celular BHK-21 / C13. Este suplemento aumentó sustancialmente las tasas de respiración mitocondrial registradas en la línea celular examinada. Debido a la estimulación de la producción de energía mitocondrial en fibroblastos normales, nuestros resultados sugirieron que Karnozin EXTRA® es un suplemento dietético potencialmente protector en la prevención de enfermedades con función mitocondrial alterada.

Role of three drugs in traumatic proliferative vitreoretinopathy in rabbits / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effect of triamcinolone acetonide(TA), artesunate(ART), and luteolin(LU)on the prevention and treatment of traumatic proliferative vitreoretinopathy(TPVR). <p>METHODS: Forty-eight cyanotic blue rabbits were selected to prepare TPVR animal models by making a penetrating eye injury and intravitreal injection of 0.3mL platelet-rich plasma, and were randomly divided into four groups(<i>n</i>=12), in which the vitreous cavity of the control group was injected with 0.1mL saline; The vitreous cavity of the TA group was injected with 0.1mL(1mg/mL)triamcinolone acetonide; The vitreous cavity of the ART group was injected with 0.1mL(20μg/mL)artesunate; 0.1mL(10μg/mL)luteolin was injected into the vitreous cavity of the LU group. The vitreous and retinal proliferation were observed by fundus photography and ocular ultrasound at 1, 2, 3 and 4wk postoperatively. The expression levels of α-SMA and VIM protein in the vitreous fluid of each group of rabbit eyes were detected by Western Blot at 28d postoperatively, and the retinal tissue structure of each group was observed by retinal HE staining. <p>RESULTS: At 28d postoperatively, the TPVR grading of rabbit eyes in the TA, ART and LU groups were significantly lower than that in the control group(<i>P</i><0.05), and the TPVR grading of rabbit eyes in the TA group was significantly lower than that in the ART and LU groups(<i>P</i><0.05). The expression levels of α-SMA and VIM proteins in the vitreous fluid of the rabbit eyes in the TA, ART and LU groups were significantly lower than those in the control group at 28d after surgery(<i>P</i><0.01). The results of HE staining showed that the arrangement of retinal layers in rabbit eyesin the control group were disordered, severely distorted or locally broken, the structure of each layer were unclear, the anterior membrane was obviously thickened, and the retina was obviously detached; The arrangement of retinal layersin rabbit eyes in the LU group were slightly distorted, inflammatory exudation was visible in front of the retina, and the retina was superficially detached; The structure of retina in rabbit eyes in the ART group were clear, with mild edema and superficial detachment; The structure of retinal layers in rabbit eyes in the TA group were clear, the arrangement was still neat, the retinal folds were locally visible, and there was no retinal detachment.<p>CONCLUSION: Intravitreal injection of triamcinolone acetonide, artesunate and luteolin were all effective in preventing and treating traumatic TPVR, among which triamcinolone acetonide has the most obvious effect.

Effects of RhoA silencing on the migration and invasion of salivary adenoid cystic carcinoma / 口腔疾病防治

Objective@# To explore whether RhoA plays a role in the migration and invasion of the salivary adenoid cystic carcinoma cell lines SACC-LM and SACC-83.@*Methods@#Total RNA and total protein were extracted from 20 salivary adenoid cystic carcinoma (SACC) and normal adjacent tissues frozen in liquid nitrogen to detect RhoA expression. RhoA-siRNA was constructed to transfect two cell lines (SACC-LM and SACC-83) for cytological experiments. The research included an experimental group (RhoA-siRNA transfection), negative control group (siRNA-NC transfection) and blank group by transient transfection with liposomes. Expression of RhoA mRNA and protein as well as the protein expression of biomarkers of epithelial-mesenchymal transition (EMT) were analyzed, including E-cadherin, N-cadherin, and Vimentin. Furthermore, the changes in invasion and migration of cells in each group were analyzed by comparing the number of transmembrane cells in the Transwell assay and the results of the scratch test.@*Results@#Compared with normal adjacent tissues, RhoA protein and mRNA levels increased in SACC tissues. Compared with the control group, the relative expression levels of RhoA mRNA and protein decreased (P < 0.01), the relative expression levels of E-cadherin protein increased, and the relative expression levels of N-cadherin and vimentin protein increased in the experimental group (P < 0.01). Additionally, the trial results revealed that RhoA knockdown restrained cell migration and invasion (P < 0.01).@*Conclusion @#RhoA expression increased in SACC tissue. Silencing RhoA in vitro could effectively restrain cell migration and invasion in SACC-LM and SACC-83 cells through the regulation of EMT signaling pathways.

Effects of EFHD2 protein deletion on tight junction proteins in mouse Sertoli cells / 安徽医科大学学报

Objective @#To study the effect of EFHD2 protein deletion in Sertoli cells on Occludin，a component of tight junction protein and the localization and expression of EF-hand domain family member D2 (EFHD2) in mouse testis．@*Methods @#Total RNA and protein were extracted from adult mice's heart ，liver ，spleen ，lung ，kidney， brain and testis tissues．The mRNA and protein levels of EFHD2 in each organ tissue were detected by qRT-PCR and Western blot．Immunofluorescence and immunohistochemistry detected the localization and expression of EF- HD2 in testicular tissues．SiRNA interference was used to reduce EFHD2 in Sertoli cells to detect Occludin protein expression. @*Results @#qRT-PCR showed that the expression of EFHD2 was the highest in the testis．Western blot results showed that the expression level increased in testis tissue．Indirect immunofluorescence and immunohisto- chemistry results showed that the protein was mainly distributed in Sertoli cells and co-localized with cytoskeletal Vimentin，indicating that the protein was expressed in Sertoli cells．After the decrease of EFHD2 protein expres- sion，Occludin protein expression also decreased．@*Conclusion @#The expression of EFHD2 protein in the testis is relatively high，mainly distributed in Sertoli cells of the testis，co-localized with Vimentin，and can affect the nor- mal expression of tight junction protein Occludin．It is suggested that EFHD2 can promote and maintain the junction structure of Sertoli cells and provide a stable microenvironment for spermatogenesis.

Overexpression of miR-34c-5p inhibits invasion and migration of gastric cancer cells / 中国药理学通报

Aim To study the expression of microRNA(miR-34c-5p)in gastric cancer tissues and the mechanism of regulating the invasion and migration of gastric cancer cells. Methods The expression of miR-34c-5p mRNA in human gastric cancer tissues and cells was detected by quantitative real-time polymerase chain reaction(RT-qPCR). An overexpression model of miR-34c-5p was constructed in human gastric cancer cells. The transfection of lentivirus was observed under inverted fluorescence microscope, and the transfection efficiency was verified by RT-qPCR. Scratch test and Transwell assay were used to detect the effects of overexpression of miR-34c-5p on invasion and migration of gastric cancer cells. Western blot was used to detect the expression of MAP2K1, E-Cadherin and Vimentin in gastric cancer cells of each group. Dual-luciferase reporter assay was employed to detect whether MAP2K1 was a target gene of miR-34c-5p. Results The expression of miR-34c-5p mRNA in 10 human gastric cancer tissues was significantly lower than that in adjacent tissues(P<0.05). The expression of miR-34c-5p mRNA in gastric cancer HGC-27 cell was significantly lower than that in normal gastric mucosa GES-1 cell(P<0.01). Compared with blank control group and miR-34c-5p-NC group, miR-34c-5pmimics group significantly increased its miR-34c-5p mRNA expression level(P<0.05). The results of scratch test and Transwell experiment showed that overexpression of miR-34c-5p significantly inhibited the migration and invasion ability of gastric cancer cells(P<0.05, P<0.01). Western blot results showed that the protein expression levels of MAP2K1 and Vimentin significantly decreased and E-Cadherin markedly increased in gastric cancer HGC-27 cells after overexpression of miR-34c-5p(P<0.05). In addition, compared with mimic-NC+miR-34c-5p-mimics group, the lucifase activity of pmiR-MAP2K1-WT plasmid decreased(P<0.01), while the lucifase activity of pmiR-MAP2K1-MUT plasmid was not changed. Conclusions The low expression of miR-34c-5p in gastric cancer tissues and cells may inhibit the invasion and migration of gastric cancer cells through targeted regulation of MAP2K1 expression.

Why human prepuce is a valuable resource in science? Histological, ultrastructural, immunohistochemical and statistical analysis / ¿Por qué el prepucio humano es un recurso valioso en la ciencia? Análisis histológicos, ultraestructurales, inmunohistoquímicos y estadísticos

SUMMARY: This research was to examine the histological and ultrastructural characteristics of prepuce samples, as well as vimentin and S100 protein localization and statistical analysis. Urologists have long struggled with the prepuce, which is used to treat a variety of urethral problems. Skin biopsies were collected from the prepuce at the moment of circumcision and processed for light microscopy, electron microscope examination, immunohistochemical techniques, and statistical analysis in a total of six boys. Histologically, the prepuce epidermis displayed focal spiky ridges, which are saw-toothed interspersed with sulci, slight hyperpigmentation, looser connective tissue and plentiful vascular components. Immunohistochemically, the existence of melanocytes and Langerhans cells in the epidermis, as well as smooth muscles in the dermis, was stained positively for vimentin. Also, there was a positive reactivity of the Langerhans cells in the epidermis and around Meissner's corpuscles in the dermis for S100 protein staining. Ultrastructurally, the prepuce's intercellular gaps were widened, melanocytes rested on a folded basement membrane, and desmosomal content was reduced, with a prominent active euchromatic nucleus. Cytoplasmic projections were distended and elongated, and the interstitial blood vessels were surrounded by endothelial cells and rested on a basement membrane. There were also minimal collagen fibers in the interstitium. The prepuce's histological and ultrastructural features, as well as immunohistological studies using vimentin and S100 protein as intermediate filaments and statistical analysis, all demonstrated that it is a useful scientific resource.

RESUMEN: El presente trabajo de investigación se realizó para examinar las características histológicas y ultraestructurales de las muestras de prepucio, así como la localización y el análisis estadístico de la vimentina y la proteína S100. Los urólogos han intentado trabajar durante mucho tiempo con el prepucio, que se usa para tratar una variedad de problemas uretrales. Se recolectaron biopsias de piel del prepucio de seis niños en el momento de la circuncisión y se procesaron para microscopía óptica, examen con microscopio electrónico, técnicas inmunohistoquímicas y análisis estadístico. Histológicamente, la epidermis del prepucio mostraba crestas puntiagudas focales, intercaladas con surcos, hiperpigmentación leve, tejido conectivo más laxo y abundantes componentes vasculares. Inmunohistoquímicamente, la existencia de melanocitos y células dendríticas epidérmicas (células de Langerhans), así como músculo liso en la dermis, se tiñeron positivamente para vimentina. Además, hubo una reactividad positiva de las células dendríticas epidérmicas en la epidermis y alrededor de los corpúsculos del tacto (de Meissner) en la dermis para la tinción de la proteína S100. Ultraestructuralmente, los espacios intercelulares del prepucio se ensancharon, los melanocitos descansaban sobre una membrana basal plegada y el contenido desmosómico se redujo, con un núcleo eucromático activo prominente. Las proyecciones citoplasmáticas estaban distendidas y alargadas, y los vasos sanguíneos intersticiales estaban rodeados por células endoteliales y descansaban sobre una membrana basal. También había fibras de colágeno mínimas en el intersticio. Las características histológicas y ultraestructurales del prepucio, así como los estudios inmunohistológicos utilizando vimentina y proteína S100 como filamentos intermedios y el análisis estadístico, demostraron que es un recurso científico útil.

Effects of Ferulic Acid on the Proliferation ，Invasion and Apoptosis of HepG 2 Hepatocellular Carcinoma Cells / 中国药房

OBJECTIVE：To study the effects of ferulic acid on t he proliferation ，invasion and apoptosis of HepG 2 hepatocelluar carcinoma cells. METHODS ：CCK-8 assay was used to screen the concentration of ferulic acid. Western blot assay was adopted to screen the optimal concentration of interleukin 6（IL-6）to induce HepG 2 cell model with high expression of phosphorylated signal transduction protein and activator 3（p-STAT3）protein. HepG 2 cell were divided into blank control group ， model group ，ferulic acid group （0.5 mmol/L）and positive control group （p-STAT3 inhibitor C 188-9，10 μmol/L）. Except for blank control group ，model group treated with IL- 6，while administration groups were treated with IL- 6 and relevant drugs. Cell survival rate ，invasion and apoptosis rate in early and late stage were detected by CCK- 8 assay，Transwell assay and Annexin V-FITC/PI double staining ，respectively. Western blot assay was used to detect the expression of p-STAT 3，caspase-3，ZBP-89 and vimentin proteins in each group. On the basis of the PDB protein database ，using 1BG1，a highly similar crystal structure of STAT3，as docking template ，using the region around Tyr 705 as the putative binding pocket ，the docking analysis of ferulic acid with STAT 3 protein was carried out. RESULTS ：It is selected to use 0.5 mmol/L ferulic acid intervention for 48 h as the follow-up experimental condition ；50 ng/mL IL- 6 was selected as the modeling condition. Compared with blank control group ，the number of cell invasion ，p-STAT3/STAT3 ratio and protein expression of vimentin were increased significantly in model group （P＜0.05 or P＜0.01），while late apoptosis rate and protein expression 20 of caspase- 3 were decreased significantly （P＜0.05 or P＜ 0.01）. Compared with model group ，cell survival rate ，the number of cell invasion ，p-STAT3/STAT3 ratio and protein expression of vimentin were d ecreased significantly in ferulic acid group and positive control group （P＜0.05 or P＜0.01）；early apoptotic rate （except for ferulic acid group ），late apoptotic rate，the protein expression of caspase- 3 and ZBP- 89（except for positive control group ）were increased significantly （P＜0.05 or P＜0.01）. The results of molecular docking showed that the carboxylic groups of ferulic acid could interact with 1.9 Å hydrogen bond of Asn 581 and 2.0 Å hydrogen bond of Lys 591，with binding energy of －4.4 kcal/mol. CONCLUSIONS ：Ferulic acid may inhibit the activity of p-STAT 3 by directly binding to the phosphorylation site of STAT 3；it may up-regulate the protein expression of caspase- 3 via STAT 3 dependent pathway ，or up-regulate the protein expression of ZBP- 89 via STAT 3 independent pathway and then down-regulate the protein expression of vimentin ，so as to inhibit the proliferation ，invasion and apoptosis of HepG 2 cells.

Diferencias fenotípicas de fibroblastos gingivales en sujetos con hiperplasia gingival idiopática frente a sujetos periodontalmente sanos: estudio piloto / Phenotypic differences of gingival fibroblasts in subjects with idiopathic gingival hyperplasia versus periodontally healthy subjects: a pilot study

Objetivo: identificar, describir y diferenciar las características fenotípicas de los fibroblastos gingivales (FGs) en pacientes con hiperplasia gingival idiopática (HGI) e individuos periodontalmente sanos. Métodos: los FGs fueron aislados a partir de tejido gingival de individuos periodontalmente sanos (n=2) y pacientes con HGI (n=2). Los FGs se cultivaron en el medio DMEM (Dulbecco's Modified of Eagle Medium) a 37°C con 5% de CO2. La identificación y localización de la actina, vimentina y mitocondrias en FGs fue realizada y evaluada microscópicamente mediante inmunofluorescencia con anticuerpos monoclonales. La capacidad de migración de los FGs en los pacientes con HGI e individuos sanos también fue estudiada. Resultados: todos los FGs fueron mononucleares, fusiformes y con prolongaciones citoplasmáticas visibles. La faloidina permitió identificar una densa red de actina en los FGs de pacientes con HGI, contrariamente a los FGs de individuos periodontalmente sanos. La vimentina y mitocondrias fueron identificadas en los FGs de individuos sanos y pacientes con HGI sin ninguna alteración en su expresión y localización. La migración de la monocapa de los FGs indicó una actividad de migración celular importante en los FGs de los pacientes con HGI, en relación a los FGs de los individuos periodontalmente sanos. Conclusión: los FGs de pacientes con HGI conservan características fenotípicas celulares similares a los FGs de individuos periodontalmente sanos. Sin embargo, los FGs de pacientes con HGI simulan tener una mayor capacidad migratoria que amerita ser explorada en futuros trabajos de investigación.

Objective: To identify and to describe the phenotypic characteristics of gingival fibroblasts from patients with idiopathic gingival hyperplasia (IGH) and periodontally healthy individuals. Methods: Gingival fibroblasts (GFs) were isolated from gingival tissue from periodontally healthy individuals (n=2) and patients with IGH (n=2). The GFs were grown in DMEM (Dulbecco's Modified of Eagle Medium) at 37°C with 5% CO2. The identification and location of actin, vimentin and mitochondria in GFs were performed and evaluated microscopically by immunofluorescence with monoclonal antibodies. The migration capacity of GFs from IGH and healthy individuals was also studied. Results: All the GFs were mononuclear, fusiform and with visible cytoplasmic extensions. The phalloidin allowed to identify a dense actin network in the GFs of patients with IGH, contrary to the GFs of periodontally healthy individuals. Vimentin and mitochondria were identified in the GFs of healthy individuals and patients with IGH without any alteration in their expression and location. Monolayer migration of GFs indicates significant cell migration activity in the GFs of patients with IGH in relation to the GFs of periodontally healthy individuals. Conclusion: GFs from patients with IGH retain cellular phenotypic characteristic similar to GFs from periodontally healthy individuals. However, the GFs of patients with IGH simulate having a greater migratory capacity that deserves to be explored in future research works.

Anti-fibrosis effects of polysaccharide of Prismatomeris tetrandra on silicosis rats / 中草药

Objective: To investigate the anti-fibrosis effects of polysaccharide of Prismatomeris tetrandra on the silicosis rats and the underlying mechanism. Methods: The experimental silicosis rat model was established by oropharyngeal aspiration method. Forty male adult Wistar rats were randomly divided into two groups: saline group and modeling group. The experimental fibrosis of silicosis was induced by dripping 1 mL of 50 mg/mL SiO2 onto the oropharynx of rats of the modeling group. After modeling, the modeling group were randomly divided into one model group with SiO2 and three polysaccharide groups with SiO2 plus low, medium, and high dose of polysaccharide of P. tetrandra, eight rats per group. One day after modeling, rats in three polysaccharide groups were administered with the polysaccharide at a dose of 125, 250, and 500 mg/kg, respectively, daily for 56 d, meanwhile, the saline group and the model group were given the same amount of saline daily for 56 d. The rats were then sacrificed at the 56th day of the experiment and lung tissues were collected. The pathological changes and fibrosis of the lung tissues were observed by HE staining and Masson staining. Real-time quantitative PCR method was used to detect the mRNA expression of epithelial calcium adhesion protein (E-cadherin) and alpha smooth muscle actin (α-SMA) in lung tissue; Western blotting method was used to measure the levels of E-cadherin, α-SMA, and Vimentin in lung tissue. Results: There were no significant differences in body weight between groups. Compared with the saline group, the alveolar inflammation and the pulmonary fibrosis were aggravated, the lung/body coefficient was increased, the levels of α-SMA mRNA and α-SMA protein were significantly increased, the mRNA and protein expression level of E-cadherin was decreased markedly, and the level of Vimentin was increased significantly in the model group. Compared with the model group, the alveolar inflammation and the pulmonary fibrosis were alleviated, the lung/body coefficient was decreased, the levels of E-cadherin mRNA and protein were increased markedly, the levels of α-SMA mRNA and protein expression were decreased sharply, and the levels of Vimentin were decreased significantly in the polysaccharide groups. Conclusion: The polysaccharide of P. tetrandra exhibited the anti-fibrosis effects on silicosis by reducing silicosis-induced lung injury and slowing down the process of pulmonary fibrosis.

Establishment of a vimentin knockout and HIV-1 gp120 transgenic mouse model / 南方医科大学学报

OBJECTIVE@#To construct a HIV-1 gp120 transgenic mice (gp120 Tg) with vimentin (VIM) gene knockout.@*METHODS@#Female HIV-1 gp120 Tg mice were mated to VIM heterozygote mice (F0). All the offspring mice were derived from these original founders so that both genotypes had the same mixed genetic background. The F1 mice were bred to generate of VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. PCR was performed for genotyping of the mice, and the expressions of VIM and gp120 in the brain tissues were examined using immunoblotting.@*RESULTS@#The results of PCR showed the presence of the target bands in VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. In VIM/gp120 Tg mice, gp120 expression was detected throughout the brain regions while no VIM expression was detected.@*CONCLUSIONS@#We generated gp120 transgenic mouse models with VIM gene knockout, which facilitate the exploration of the role of VIM in gp120-induced neurotoxicity.

The role of transforming growth factor β1 and collagen in vulvar lichen sclerosis / 医学研究生学报

ObjectiveTo investigate the expression of transforming growth factor β1 (TGF-β1) and collagen I, III (Col I, III) in vulvar lichen sclerosis (VLS) and their role in VLS.MethodsThe specimens of30 VLS tissues (15 in early stage and 15 in progressive stage), and 15 vulvar normal skin tissues were selected by biopsy or surgical excision from 2016 to 2018 in our hospital. The expression of Vimentin was detected by immunohistochemistry. The expression of TGF-β1, TGF-β2, Col I and Col III mRNA was detected through RT-PCR. Simultaneously, the fibroblasts were visualized in diseased tissues by labeling Vimentin.ResultsThe expression of Vimentin in VLS was increased significantly (P0.05). Col I mRNA was up-regulated in VLS, obviously in early stage. Meanwhile, Col III mRNA down-regulated gradually from the early to the progressive stage of VLS. Therefore, the Col I/III ratio increased gradually.ConclusionThe increase of fibroblasts and TGF-β1 in dermis of VLS promotes the synthesis of Col I and reduces the content of Col III, which may be one of the factors leading to the decrease of skin elasticity in VLS.