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Objective:To analyze the value of laminin γ2 (LAMC2) in the diagnosis of hepatocellular carcinoma (HCC) and the difference in patients with different types of microvascular invasion (MVI).Methods:A cohort of 100 patients with HCC who underwent surgical treatment at the Faculty of Hepato-Pancreato-Biliary Surgery, Chinese PLA General Hospital from January 2021 to March 2022 were prospectively enrolled. There were 80 males and 20 females, aged (55.7±11.1) years. The data of 17 patients with hepatic hemangioma without cirrhosis who underwent operation at the same hospital during the study period were collected to serve as the control group (6 males, 11 females), aged (42.8±9.8) years. LAMC2 in serum was determined by enzyme linked immunosorbent assay. The levels of alpha-fetoprotein (AFP) and LAMC2 were compared between the two groups, and receiver operating characteristic (ROC) curves were drawn to compare these two markers in the diagnosis of HCC. The LAMC2 of different MVI patients were compared.Results:The levels of LAMC2 and AFP were 1 334.2(838.9, 2 656.0) pg/ml and 19.0(4.6, 778.6) μg/L in the HCC group, which were significantly higher than 375.2(221.2, 691.7)pg/ml and 3.3(2.5, 3.5) μg/L in the control group ( Z=-4.32, -4.63, both P<0.001). The areas under the ROC curve were 0.829(95% CI: 0.748-0.892) for LAMC2 and 0.852(95% CI: 0.769-0.910) for AFP, and was 0.949(95% CI: 0.911-0.988) for using both in the diagnoses. The diagnostic efficacy of combining LAMC2 and AFP was significantly better than that of LAMC2 alone and AFP alone (area under ROC: Z=3.15, 3.07, P=0.002, 0.002). When the patients were divided into the M0 group (61 patients), the M1 Group (25 patients) and the M2 Group (14 patients) based on MVIs, the concentrations of LAMC2 were 1 168.6(834.3, 2 521.4) pg/ml, 942.2(614.0, 2 056.6) pg/ml and 3 128.4(1 852.7, 7 191.3) pg/ml, respectively. The level of LAMC2 in the M2 group was significantly higher than that in the M0 and M1 groups ( Z=-3.46, -3.32, P=0.001, 0.004). Conclusion:The diagnostic efficacy of LAMC2 combined with AFP for HCC was significantly higher than that of either LAMC2 alone or AFP alone. Serum LAMC2 levels were significant different among the groups of HCC patients with different types of MVI.
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Laminin subunit alpha 4 (LAMA4),a member of the laminin family,is present in the intercellular matrix of adult tissues as a major component of basement membrane.LAMA4 is involved in the adhesion of cells and can bind to corresponding integrins to activate relevant signaling pathways,playing an essential role in the growth,proliferation,and migration of cells.It has been demonstrated that LAMA4 is associated with the occurrence and development of a variety of diseases including tumors,and the expression of LAMA4 can be used as a biomarker of tumor diagnosis and prognosis.This paper summarizes the current research progress in LAMA4 with the focus on the relationship between LAMA4 and diseases,especially tumor,with a view to provide new directions for the future research.
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Adult , Humans , Laminin , Extracellular MatrixABSTRACT
A Distrofia muscular laminin subunit alpha 2 (DM LAMA2) é caracterizada pela deficiência da proteína da cadeia laminina α2, apresentando sintomas distróficos que progridem na infância. Objetivo: Apresentar dados da avaliação fisioterapêutica de uma criança com DM LAMA2 em acompanhamento ambulatorial. Métodos: Pesquisou-se em prontuário dados referentes às avaliações fisioterapêuticas de uma criança de 12 anos com diagnóstico de DM LAMA2 atendida em ambulatório especializado. A avaliação caracterizou-se por ausculta pulmonar, espirometria, verificação de parâmetros cardiorrespiratórios, análise do pico de fluxo de tosse (PFT), força muscular respiratória (FMR) e avaliação motora através da escala MFM-32. Resultados: Realizou-se três avaliações durante 9 meses. A paciente possui doença pulmonar restritiva, na primeira avaliação apresentou VEF1= 29% na espirometria e, comparando com a terceira avaliação obteve aumento de 1%, também houve aumento de 2% na relação VEF1/CVF, 5% no PEF e 11% no FEF25-75%. Na FMR, obteve-se valores de PImáx= 17,9% e PEmáx= 7,13% na primeira avaliação com aumento de 16,85% e 5,34% respectivamente, entre primeira e terceira avaliação. O PFT manteve-se em 0L/min em todas avaliações. Na primeira avaliação motora pontuou 25% no escore total da escala MFM-32, aumentando 3,12% na terceira avaliação. Ao longo do acompanhamento iniciou-se o uso da VNI noturna e introduzida a técnica de AS, buscando correção da hipoxemia, apneias noturnas, aumento do PFT e FMR. Conclusão: Paciente apresentou manutenção de parâmetros espirométricos, aumento de variáveis de FMR e função motora, sem piora do quadro. Possíveis resultados decorridos do acompanhamento multiprofissional e especializado.
Laminin subunit alpha 2 muscular dystrophy (DMLAMA2) is characterized by a deficiency of the α2 laminin chain protein, presenting dystrophic symptoms that progress in childhood. Objective: To present data from the physical therapy evaluation of a child with DMLAMA2 in outpatient follow-up. Methods: Medical records were searched for data referring to the physiotherapeutic evaluations of a 12-year-old child diagnosed with DMLAMA2 treated at a specialized outpatient clinic. The evaluation was characterized by pulmonary auscultation, spirometry, verification of cardiorespiratory parameters, analysis of peak cough flow (PFT), respiratory muscle strength (FMR) and motor evaluation using the MFM-32 scale. Results: Three evaluations were carried out during 9 months. The patient has restrictive disease, in the first evaluation she had FEV1= 29% in spirometry, compared to the third evaluation, she had an increase of 1%, there was also an increase of 2% in the FEV1/FVC ratio, 5% in the PEF and 11% in the FEF25-75%. In the FMR, values of MIP=17.9% and MEP= 7.13% were obtained in the first assessment, with an increase of 16.85% and 5.34%, respectively, between the first and third assessments. The PFT remained at 0L/min in all evaluations. In the first motor evaluation, it scored 25% in the total score of the MFM-32 scale, increasing 3.12% in the third evaluation. During the follow-up, the use of nocturnal NIV started and AS technique was introduced, seeking correction of hypoxemia, night apneas, increased PFT and FMR. Conclusion: Patient presented maintenance of spirometric parameters, increased FMR variables and motor function, without worsening the condition. Possible results from the multidisciplinary and specialized monitoring.
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Objective To evaluate the performance of FibroTouch in combination with four hepatic fibrosis biomarkers for assessment of the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver disorders. Methods A total of 63 patients with chronic schistosomiasis-induced liver diseases admitted to The Third People’s Hospital of Kunshan City from January to March 2021 were enrolled as the observation group, while 50 healthy volunteers receiving health examinations in the hospital during the study period were randomly selected as the control group. The liver stiffness measurement (LSM) was determined using the FibroTouch technique, and the serum levels of four hepatic fibrosis biomarkers were detected using chemilumi-nescence immunoassay, including type IV collagen (IV-C), type III procollagen (PC-III), hyaluronidase (HA) and laminin (LN). The receiver operating characteristic (ROC) curves of LSM and four hepatic fibrosis biomarkers alone and in combination for assessing the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver disorders were plotted and the area under the ROC curve (AUC) was estimated to examine the value of LSM and four hepatic fibrosis biomarkers alone and in combination for assessing the degree of hepatic fibrosis. Results There were 63 subjects in the observation group, including 28 men and 35 women, and the participants had a mean age of (65.34 ± 12.56) years and a mean body mass index (BMI) of (24.47 ± 11.05) kg/m2. There were 50 subjects in the control group, including 22 men and 28 women, and the participants had a mean age of (64.28 ± 13.10) years and a mean BMI of (25.12 ± 11.64) kg/m2. There were no significant differences between the observation and control groups in terms of gender ratio (χ2 = 0.002, P > 0.05), age (t = 0.437, P > 0.05) or BMI (t = 0.303, P > 0.05). The LSM [(8.65 ± 5.22) vs. (3.24 ± 1.10) kPa; t = 8.013, P < 0.05], IV-C [(51.80 ± 9.45) vs. (30.10 ± 10.34) ng/L; t = 11.506, P < 0.05], PC-III [(77.28 ± 17.22) vs. (48.62 ± 9.54) ng/L; t = 11.224, P < 0.05], HA [(39.55 ± 5.32) vs. (84.89 ± 10.34) ng/L; t = 30.158, P < 0.05] and LN [(99.47 ± 7.37) vs. (61.93 ± 9.80) ng/L; t = 22.496, P < 0.05] were significantly greater in the observation group than in the control group, and Spearman correlation analysis showed that the degree of liver fibrosis positively correlated with LSM (rs = 0.675, P < 0.01), IV-C (rs = 0.421, P < 0.01), PC-III (rs = 0.517, P < 0.01), HA (rs = 0.550, P < 0.01) and LN (rs = 0.539, P < 0.01) among patients with chronic schistosomiasis-induced liver diseases. ROC curve analysis revealed that the AUC of LSM for assessment of the hepatic fibrosis degree was 0.884 (P < 0.001), and the LSM cutoff, sensitivity and specificity were 11.75 kPa, 71.43% and 84.00% at the highest Youden index, respectively. In addition, the AUC of four hepatic fibrosis biomarkers for assessment of the hepatic fibrosis degree was 0.577 to 0.670, with 70.174 to 115.237 ng/L cutoff values, 17.46% to 68.25% sensitivity and 71.01% to 96.00% specificity. In addition, the sensitivity and specificity of LSM combined with four hepatic fibrosis biomarkers were 92.06% and 95.07% for assessment of the hepatic fibrosis degree among patients with chronic schistosomiasis-induced liver diseases. Conclusion FibroTouch in combination with detection of four hepatic fibrosis biomarkers has a high sensitivity and specificity for assessing the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver diseases, which deserves widespread clinical uses.
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Objective:To analyze clinical and immunoserological features of patients with anti-p200 pemphigoid.Methods:Clinical data were collected from patients with confirmed anti-p200 pemphigoid in Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2015 to October 2021, and their clinical and immunoserological characteristics were retrospectively analyzed.Results:Seven patients with anti-p200 pemphigoid were included. Indirect immunofluorescence on salt-split skin (IIF-SSS) showed that serum IgG antibodies of the 7 patients were located in the dermis of the salt-split skin, and Western blot analysis with dermal extracts as substrates revealed a protein band with a relative molecular mass of 200 000. Four patients presented with classic bullous pemphigoid-like skin lesions, 2 initially presented with eczematous lesions, and 1 presented with linear IgA bullous dermatosis-like skin lesions. Circulating IgG antibodies could recognize the recombinant laminin γ1 C-terminal region in 6 cases. Four patients received different doses of systemic glucocorticoids, 1 of whom was resistant to high-dose systemic glucocorticoids (equivalent to 1.4 mg·kg -1·d -1 prednisone) ; 2 responded well to minocycline and dapsone; 1 was lost to follow-up. Four patients achieved complete remission and discontinued the treatment at a mean follow-up of 22.5 months; 2 received complete remissiona on minimal therapy at a mean follow-up of 8 months. Conclusion:Patients with anti-p200 pemphigoid presented with heterogeneous clinical manifestations, and the recombinant C-terminal fragment of laminin γ1 can serve as a reliable antigen substrate for the detection of autoantibodies in patients with anti-p200 pemphigoid; some patients can eventually achieve complete remission off treatment.
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BACKGROUND Clostridioides difficile is the most common cause of nosocomial diarrhea associated with antibiotic use. The disease's symptoms are caused by enterotoxins, but other surface adhesion factors also play a role in the pathogenesis. These adhesins will bind to components of extracellular matrix. OBJECTIVE There is a lack of knowledge on MSCRAMM, this work set-out to determine the adhesive properties of several C. difficile ribotypes (027, 133, 135, 014, 012) towards laminin-1 (LMN-1). METHODS A binding experiment revealed that different ribotypes have distinct adhesion capabilities. To identify this adhesin, an affinity chromatography column containing LMN-1 was prepared and total protein extracts were analysed using mass spectrometry. FINDINGS Strains from ribotypes 012 and 027 had the best adhesion when incubated with glucose supplementations (0.2%, 0.5%, and 1%), while RT135 had a poor adherence. The criteria were not met by RT014 and RT133. In the absence of glucose, there was no adhesion for any ribotype, implying that glucose is required and plays a significant role in adhesion. MAIN CONCLUSIONS These findings show that in the presence of glucose, each C. difficile ribotype interacts differently with LMN-1, and the adhesin responsible for recognition could be SlpA protein.
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Abstract Potent signaling agents stimulate and guide pulp tissue regeneration, especially in endodontic treatment of teeth with incomplete root formation. Objective This study evaluated the bioactive properties of low concentrations of extracellular matrix proteins on human apical papilla cells (hAPCs). Methodology Different concentrations (1, 5, and 10 µg/mL) of fibronectin (FN), laminin (LM), and type I collagen (COL) were applied to the bottom of non-treated wells of sterilized 96-well plates. Non-treated and pre-treated wells were used as negative (NC) and positive (PC) controls. After seeding the hAPCs (5×103 cells/well) on the different substrates, we assessed the following parameters: adhesion, proliferation, spreading, total collagen/type I collagen synthesis and gene expression (ITGA5, ITGAV, COL1A1, COL3A1) (ANOVA/Tukey; α=0.05). Results We observed greater attachment potential for cells on the FN substrate, with the effect depending on concentration. Concentrations of 5 and 10 µg/mL of FN yielded the highest cell proliferation, spreading and collagen synthesis values with 10 µg/mL concentration increasing the ITGA5, ITGAV, and COL1A1 expression compared with PC. LM (5 and 10 µg/mL) showed higher bioactivity values than NC, but those were lower than PC, and COL showed no bioactivity at all. Conclusion We conclude that FN at 10 µg/mL concentration exerted the most intense bioactive effects on hAPCs.
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Humans , Extracellular Matrix Proteins , Fibronectins , Cell Adhesion , Cells, Cultured , Laminin , Collagen Type I , Extracellular MatrixABSTRACT
Objective:To investigate the correlations of laminin subunit gamma 3 (LAMC3) expression with prognosis of ovarian cancer (OC).Methods:LAMC3 protein expression was measured using immunohistochemical streptavidin-peroxidase-biotin connection method (IHC). Gene expression and related clinical data in the cancer genome atlas (TCGA) cohort and clinical proteomic tumor analysis consortium (CPTAC) were applied to analyse the correlation between gene and protein expressions and clinical outcomes. Correlations between LAMC3 and clinicopathological factors were evaluated using the Pearson χ2 test (2-sided). The probability of survival and significance was calculated using the Kaplan-Meier plot. The functional clustering of biological pathways enriched from co-expressed genes of LAMC3 was used to explore the possible mechanisms that LAMC3 might contribute to poor prognosis. Results:Based on the IHC results of 216 OC tissues or ovaries (including 208 tumors and 8 normal tissues) and 51 OC tissues (including 24 chemotherapy-resistant and 27 sensitive tissues), and the protein expression data from CPTAC (including 100 primary tumors and 25 normal tissues), the results showed that the protein expression of LAMC3 was significantly decreased in OC tissues compared with normal, decreased in advanced-stage tissues compared with early-stage tissues, and decreased in drug-resistant tissues compared with sensitive tissues (all P<0.05). Furthermore, low expression of LAMC3 protein was significantly associated with poor disease-free survival (DFS) and overall survival (OS) in 51 OC tissues ( P<0.01), consistent with the results that the low levels of LAMC3 mRNA predicted short DFS and OS in 489 OC tissues of the TCGA cohort ( P<0.05). The results suggested that low expression of LAMC3 might be the adverse factors for OC development, such as drug resistance and advanced tumors, and might be a risk indicator for prognosis. Moreover, functional clustering of biological pathways enriched from the co-expressed genes of LAMC3 in TCGA ovarian cohort indicated that LAMC3 potentially involved in regulation of OC via oncogene-pathways such as Ras associated protein 1 (Rap1), mitogen-activated protein kinase (MAPK), Ras and cell adhesion-related pathways such as extra cellular matrix (ECM)-receptor interaction and focal adhesion. It indicated that LAMC3 might contribute to short survival and tumor progression by regulation of the above pathways. Conclusion:Low expression of LAMC3 is related to poor prognosis and malignant progression in OC, and thus it is expected to be a new prognostic marker and therapeutic target for clinical treatment.
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SUMMARY: The aim of this study was to evaluate the effects of stretching and therapeutic ultrasound (TUS) on desmin and laminin contents of rat muscle after contusion. Male Wistar rats (n = 35, 8-9 weeks of age, 271 ± 14g body weight) were divided into five groups: Control group (CG) (n= 03); Injured group (IG) (n= 8); Injured + ultrasound group (IUSG) (n= 8); Injured+stretching group (ISG) (n= 8); Injured +ultrasound + stretching group (IUSSG) (n= 8). The application of ultrasound started 72 hours after the contusion, using the 50 % pulsed mode, 0.5 W/cm2, 5 min, once a day, for five consecutive days. Passive manual stretching was started on the tenth day after injury, with four repetitions of 30 s each and 30 s rest between repetitions, once a day, five times per week, for a total of ten applications. After 22 days, the rats were euthanazied and the gastrocnemius of both limbs removed for desmin and laminin immunohistochemistry morphometric measurement. Analysis was conducted using ANOVA one way post-hoc Tukey to parametric data and Kruskall-Wallis for non-parametric data. The IUSSG animals showed a larger area of desmin than ISG (p<0.05). It was found a decrease in laminin comparing IUSG to IG. However, laminin area was higher in ISG than all groups (p<0.05). UST isolated or in combination with stretching influenced gastrocnemius regeneration in different manners. While stretching applied isolated enhanced gastrocnemius regeneration noticed by the increase in laminin area, in combination with TUS strengthened the muscle healing rising desmin area.
RESUMEN: El objetivo de este estudio fue evaluar los efectos del estiramiento y la ecografía en los contenidos de desmina y laminina del músculo de rata después de la lesión. Ratas Wistar macho (n = 35, 8-9 semanas de edad, 271 ± 14 g de peso corporal) se dividieron en cinco grupos: grupo de control (CG) (n = 03); Grupo lesionado (GL) (n = 8); Lesionado + grupo de ultrasonido (LGU) (n= 8); Lesionado + grupo de estiramiento (LGE) (n = 8); Lesionado + ultrasonido + grupo de estiramiento (LUGE) (n = 8). La aplicación de ultrasonido comenzó 72 horas después de la lesión, usando el modo pulsado al 50 %, 0,5W / cm2, 5 min, una vez al día, durante cinco días consecutivos. El estiramiento manual pasivo se inició el décimo día después de la lesión, con cuatro repeticiones de 30 seg cada una y 30 seg de descanso entre repeticiones, una vez al día, cinco veces por semana, para un total de diez aplicaciones. Las ratas fueron sacrificadas después de 22 días, y se extrajo el músculo gastrocnemio de ambos miembros para la medición morfométrica de desmina y laminina a través de inmunohistoquímica. El análisis se realizó utilizando ANOVA unidireccional Tukey post-hoc para datos paramétricos y Kruskall-Wallis para datos no paramétricos. Los animales LUGE mostraron un área mayor de desmina que LGE (p <0,05). Se encontró una disminución en la laminina comparando LGU con GL. Sin embargo, el área de laminina fue mayor en LGE que en todos los grupos (p <0,05). El tratamiento con ultrasonido aislado o en combinación con estiramiento influyó en la regeneración del músculo gastrocnemio de diferentes maneras. Si bien el estiramiento aplicado, en combinación con tratamiento de ultrasonido, fortaleció el área de desmina, la regeneración del músculo gastrocnemio mejoró por el aumento en el área de laminina aumentando la curación muscular.
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Animals , Male , Rats , Ultrasonic Therapy/methods , Muscle, Skeletal/pathology , Contusions/therapy , Muscle Stretching Exercises/methods , Immunohistochemistry , Analysis of Variance , Laminin/analysis , Rats, Wistar , Muscle, Skeletal/injuries , Desmin/analysisABSTRACT
Human placental extract has wound healing potential. Immuno-blots revealed presence of laminin in placentalextract (70 ± 0.257 lg/ml; n=3). It was purified using immuno-affinity chromatography. SDS-PAGE and SEHPLC indicated a188 kDa protein with some small peptides. Since placental laminin existed in its truncatedform, its roles in cellular migration, differentiation and wound healing were verified. Induction of cellularmigration and motility in rat fibroblasts were enhanced by placental laminin as observed from scratch woundassay. Promotion of neuronal differentiation of PC12 cells by placental laminin was observed by phase contrastmicroscopy. Confocal images showed presence of laminin on the cell surface and along the axonal processes.Significant interaction between integrin receptors and laminin responsible for cellular differentiation wasdemonstrated from co-localization experiments. Union between integrin receptor and its synthetic antagonistrevealed retarded pattern of neurite outgrowth in laminin treated cells. Animal model studies revealed fasterwound healing in the presence of placental laminin. Induction of re-epithelialization and angiogenesis inwound area by cellular proliferation and adhesion were observed. The cytokine levels showed an initial riseand gradual fall over the duration of wound healing on application of the fragmented laminin. Thus, roles ofplacental laminin in neuronal differentiation and wound healing were indicated.
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OBJECTIVE: With the emerging significance of genetic profiles in the management of endometrial cancer, the identification of tumor-driving genes with prognostic value is a pressing need. The LAMC1 gene, encoding the laminin subunit gamma 1 (LAMC1) protein, has been reported to be involved in the progression of various malignant tumors. In this study, we aimed to investigate the role of LAMC1 in endometrial cancer and elucidate the underlying mechanism.METHODS: We evaluated the immunohistochemical expression of LAMC1 in atypical endometrial hyperplasia and endometrial cancer. Within the endometrial cancer cases, we analyzed the association of LAMC1 overexpression with clinicopathological factors and prognosis. Furthermore, to indentify genes influenced by LAMC1 overexpression, we transfected HEC50B and SPAC-S cells with siRNA targeting LAMC1 and conducted microarray gene expression assays.RESULTS: While none of the atypical endometrial hyperplasia specimens exhibited LAMC1 overexpression, endometrial cancer possessed a significantly higher LAMC1 overexpression rate. LAMC1 overexpression was strongly associated with histological type, lymphovascular space invasion, lymph node metastasis, advanced International Federation of Gynecology and Obstetrics stage, and poor overall survival in endometrial cancer. Gene expression microarray analysis identified 8 genes correlated with tumor progression (LZTFL1, TAPT1, SEL1L, PAQR6, NME7, TMEM109, CCDC58, and ANKRD40) that were commonly influenced in HEC50B and SPAC-S by LAMC1 silencing.CONCLUSION: LAMC1 overexpression is a potent biomarker for identifying endometrial cancer patients needing aggressive adjuvant therapy. We elucidated 8 candidate genes that may mediate progression of LAMC1 overexpressing cancer. Further investigation of the underlying mechanism should lead to the discovery of new therapeutic targets.
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Female , Humans , Endometrial Hyperplasia , Endometrial Neoplasms , Gene Expression , Gene Expression Profiling , Gynecology , Laminin , Lymph Nodes , Microarray Analysis , Neoplasm Metastasis , Obstetrics , Prognosis , RNA, Small InterferingABSTRACT
BACKGROUND: The characteristics of laminin that can promote the proliferation of stem cells have been widely concerned. OBJECTIVE: To review the interactions between laminin and many different stem cells, and provide reliable theoretical basis for chondrogenic research and application of stem cells. METHODS: Wanfang, CNKI, PubMed and Web of Science databases were searched for articles related to mechanism of laminin, changes in stem cell behaviors, and cartilage regeneration published from January 2010 to October 2019. The retrieval terms were “laminin” and “steam cells” in Chinese and English. Duplicated and poorly related articles were excluded, and finally 57 articles were included for review. RESULTS AND CONCLUSION: (1) The structural characteristics of laminin were summarized. The spatiotemporal changes of laminin during cartilage development and degradation were analyzed. At the same time, the distribution of laminin expression in natural cartilage tissue and tissue engineered cartilage tissue was compared. (2) The effects of laminin on the proliferation of various stem cells, including embryonic stem cells, induced pluripotent stem cells and adult stem cells, were described. (3) The possible hotspots on the combination of laminin and stem cells for cartilage regeneration were proposed, with the attempt of providing theoretical basis for cartilage repair and regeneration in the future.
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Background: In recent years, high-throughput omics technologies have been widely used globally to identify potential biomarkers and therapeutic targets in various cancers. However, apart from large consortiums such as The Cancer Genome Atlas, limited attempts have been made to mine existing datasets pertaining to cancers. Methods and Results: In the current study, we used an omics data analysis approach wherein publicly available protein expression data were integrated to identify functionally important proteins that revealed consistent dysregulated expression in head and neck squamous cell carcinomas. Our analysis revealed members of the integrin family of proteins to be consistently altered in expression across disparate datasets. Additionally, through association evidence and network analysis, we also identified members of the laminin family to be significantly altered in head and neck cancers. Members of both integrin and laminin families are known to be involved in cell-extracellular matrix adhesion and have been implicated in tumor metastatic processes in several cancers. To this end, we carried out immunohistochemical analyses to validate the findings in a cohort (n = 50) of oral cancer cases. Laminin-111 expression (composed of LAMA1, LAMB1, and LAMC1) was found to correlate with cell differentiation in oral cancer, showing a gradual decrease from well differentiated to poorly differentiated cases. Conclusion: This study serves as a proof-of-principle for the mining of multiple omics datasets coupled with selection of functionally important group of molecules to provide novel insights into tumorigenesis and cancer progression.
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Objective@#To investigate the clinical value of biochemical indicator and hepatic fibrosis in differential diagnosis of patients with cirrhosis and chronic liver failure.@*Methods@#From December 2015 to December 2018, 30 patients with cirrhosis and 30 patients with chronic liver failure in Zhoushan Hospital were selected.The serum levels of ALT, AST, TBil, hyaluronic acid (HA), laminin (LN), PC-Ⅲ, collagen type Ⅳ(Ⅳ-C) were detected.@*Results@#The serum levels of ALT, AST and TBil in patients with cirrhosis were significantly lower than those in patients with chronic liver failure[(258.17±88.19)U/L vs.(818.37±375.83)U/L; (256.57±97.21)U/L vs.(738.63±329.68)U/L; (51.37±22.13)μmol/L vs.(157.27±85.60)μmol/L, t=-7.95, -7.68, -6.56, all P<0.01]. The serum levels of HA, LN, PC-Ⅲ and Ⅳ-C in patients with cirrhosis were significantly higher than those in patients with chronic liver failure[(433.10±214.39)μg/L vs.(81.56±27.80)μg/L; (157.67±52.60)μg/L vs.(66.06±24.06)μg/L; (126.63±35.24)μg/L vs.(70.96±19.71)μg/L; (162.26±53.93)μg/L vs.(76.13±27.90)μg/L, t=8.90, 8.67, 7.49, 7.76, all P<0.01]. When patients with chronic liver failure were set as state variable, AUC of ALT(0.973) was the highest in ALT, AST, TBil.When patients with cirrhosis were set as state variable, AUC of HA(0.993) was the highest in HA, LN, PC-Ⅲ and Ⅳ-C.@*Conclusion@#ALT and HA are suitable for differential diagnosis of patients with cirrhosis and chronic liver failure.
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Objective To explore the mechanism of improving angiogenesis of hepatic fibrosis by Gexia Zhuyu Decoction (GZD) through the regulation of the mRNA expression of VEGF mediated by HIF-1α. Methods A total of 108 Wistar rats were randomly divided into normal group (n = 18), model group (n = 18), N-acetylcysteine (NAC) group (n = 18), high-dose GZD group (GD, n = 18), middle-dose GZD group (GZ, n = 18), and low-dose GZD group (GX, n = 18). Hepatic fibrosis model was established by intraperitoneal injection of 50% CCl4-olive oil solution (1 mL/kg) twice a week for nine weeks. Each group was administered while model established, until the rats were sacrificed. Normal group and model group were ig given sterile water 10 mL/(kg•d), NAC group was ig given NAC 0.1 g/(kg•d), GD, GZ, GX groups were given 26, 7.8, and 3.9 g/(kg•d) GZD by oral gavage. At 3, 6, and 9 weeks, rats in the corresponding groups were randomly sacrificed. Masson staining was used to make pathological specimens, immunohistochemical analysis of Col-IV and laminin was also performed, and real-time PCR was used to detect the mRNA expression of HIF-1α and VEGF. Western blotting was used to detect the protein expression levels of VEGF and VEGFR2. Results Compared with model group, NAC group and GD group significantly inhibited the expression of LN in the extracellular matrix at 9 weeks (P<0.05). Both NAC and GD groups significantly inhibited the expression of extracellular matrix Col-IV, especially at 6 weeks and 9 weeks. NAC group, GD group, and GZ group can significantly inhibit the high expression of HIF-1α in liver tissue of rats with liver fibrosis (P<0.05). At 6 weeks and 9 weeks of administration, NAC and GD groups significantly inhibited the high expression of VEGF mRNA in liver tissue (P<0.05). Both GZD and NAC could inhibit the protein expression of VEGF and VEGFR2 in liver tissue. Conclusion GZD can regulate the expression of VEGF mRNA mediated by HIF-1α, which may be one of the key mechanisms of its anti-angiogenesis for hepatic fibrosis.
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Objective To explore the prognostic role of laminin (LN ) as a tumor biological marker in predicting the recurrence of hepatocellular carcinoma (HCC) related with HBV infection after liver transplantation (LT ) .Methods Tissue samples from 251 HBV-related HCC patients undergoing LT were immunohistochemically stained with anti-LN antibody .The relevant prognostic factors were analyzed using Spearman's rank test , Kaplan-Meier method , log-rank test and multivariate step-wise Cox regression analysis .Results The expressions of LN in tumor tissues were significantly positively correlated with tumor number (P=0 .00) ,microsatellite (P=0 .02) ,venous invasion (P=0 .048) ,pTNM tumor stage (P=0 .00) ,pre-LT serum α-fetoprotein (AFP) level (P=0 .00) ,HBV DNA level (P= 0 .02) ,HBeAg level (P= 0 .02) and tumor recurrence (P= 0 .00) respectively .Significant differences existed in 1/3/5-year overall survival or tumor recurrence-free survival rate post-LT among LN different expression (-,+ ,≥ + + ) in HBV-related HCC patients (P< 0 .05 ) . Multivariate analysis indicated that LN was a significantly independent predictor in predicting poor tumor recurrence-free survival post-LT (P=0 .01) .Conclusions LN may be a feasible marker in predicting HCC recurrence post-LT for HBV-related HCC patients .
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Pigment epithelium-derived factor(PEDF)is an endogenously produced protein widely expressed throughout the human body,which exhibits multiple and varied biological actions.Over the past few years,PEDF has been described as a multifaceted protein with anti-angiogenic, neurotrophic, neuroprotective, and anti-permeability.PEDF effectively inhibits vascular endothelial growth factor(VEGF)-driven angiogenesis and vascular permeability by regulating the intracellular proteolysis of VEGF receptors in re -cent reports.Moreover,PEDF can inhibit the occurrence of neural degenerative diseases through a variety of signaling pathways.The aim of the present review is to discuss the varied biological actions of PEDF and to identify the possible molecular mechanism underly -ing its effects.
ABSTRACT
Objective To investigate the correlation between serum laminin (LN),type collagen (ⅣC),type procollagen N-terminal peptide (PⅢNP),hyaluronic acid (HA) and the severity as well as the prognosis of idiopathic pulmonary fibrosis (IPF).Methods From February 2015 to July 2016,160 IPF patients and 160 healthy subjects as controls were enrolled in this retrospective study.Serum LN,ⅣC,PⅢNP,and HA were analyzed in IPF patients and healthy controls.Pulmonary function test and chest high resolution computed tomography (HRCT) were carried out in IPF patients.Demographics and clinical characteristics,the percentage of forced vital capacity in the prediction value (FVC％pred),the percentage of diffusing capacity of the lung for carbon monoxide in the prediction value (DLCO％pred),and HRCTscore were collected.IPF patients were followed up.Results (1)There were no significant difference between two groups in age and sex ratio.The proportion of smoker in IPF patients was significantly higher than that in healthy controls(P ＜ 0.01).(2)Serum LN(P ＜ 0.01),ⅣC(P ＜ 0.01),PⅢNP(P ＜ 0.01),and HA(P ＜ 0.01) were significantly increased in the patients with IPF compared with the healthy controls.(3)Serum LN,ⅣC,PⅢNP,and HA of IPF patients positively correlated with HRCT score,all P ＜ 0.01,and negatively correlated with FVC％pred and DLCO％pred (all P ＜ 0.05).(4)Serum LN(P ＜ 0.01),ⅣC(P ＜ 0.05),PⅢNP(P ＜ 0.01),and HA(P ＜ 0.01) in acute exacerbation IPF patients were significantly higher than those in the stable IPF patients.(5)Serum LN(P ＜ 0.01),ⅣC(P ＜0.01),PⅢNP(P ＜ 0.01),and HA(P ＜ 0.01) in the survived patients were significantly lower than those inthe dead patients.Conclusions Serum LN,ⅣC,PⅢNP,and HA may reflect IPF prognosis and the severity of IPF.
ABSTRACT
Urine-derived stem cells (USCs) are considered as a promising cell source capable of neuronal differentiation. In addition, specific growth factors and extracellular matrix are essential for enhancing their neuronal differentiation efficiency. In this study, we investigated the possibility of neuronal differentiation of USCs and the role of laminin and platelet-derived growth factor BB (PDGF-BB) as promoting factors. USCs were isolated from fresh urine of healthy donors. Cultured USCs were adherent to the plate and their morphology was similar to the cobblestone. In addition, they showed chromosome stability, rapid proliferation rate, colony forming capacity, and mesenchymal stem cell characteristics. For inducing the neuronal differentiation, USCs were cultured for 14 days in neuronal differentiation media supplemented with/without laminin and/or PDGF-BB. To identify the expression of neuronal markers, RT-PCR, flow cytometry analysis and immunocytochemistry were used. After neuronal induction, the cells showed neuron-like morphological change and high expression level of neuronal markers. In addition, laminin and PDGF-BB respectively promoted the neuronal differentiation of USCs and the combination of laminin and PDGF-BB showed a synergistic effect for the neuronal differentiation of USCs. In conclusion, USCs are noteworthy cell source in the field of neuronal regeneration and laminin and PDGF-BB promote their neuronal differentiation efficiency.
Subject(s)
Humans , Chromosomal Instability , Extracellular Matrix , Flow Cytometry , Immunohistochemistry , Intercellular Signaling Peptides and Proteins , Laminin , Mesenchymal Stem Cells , Neurons , Platelet-Derived Growth Factor , Regeneration , Stem Cells , Tissue DonorsABSTRACT
O diabetes mellitus do tipo 1 (DM1) é uma doença causada pela destruição autoimune das células-ß produtoras de insulina do pâncreas. O transplante de ilhotas pancreáticas é um procedimento tecnicamente simples sendo uma alternativa terapêutica interessante para o DM1. Entretanto, a oferta limitada de pâncreas de doadores falecidos e a necessidade de imunossupressão crônica são fatores que limitam a aplicabilidade dessa modalidade de transplante. Neste trabalho foram estudadas duas estratégias que visam oferecer soluções aos fatores limitantes do transplante de ilhotas pancreáticas. Na primeira parte do trabalho, o mecanismo molecular que dirige o processo de diferenciação de células-tronco embrionárias murinas (murine embryonic stem cells, mESCs) em células produtoras de insulina (insulin producing cells, IPCs) foi analisado visando otimizar o processo de diferenciação. Nós selecionamos o gene Thioredoxin interacting protein (Txnip), diferencialmente expresso ao longo da diferenciação ß-pancreática, para realizar um estudo funcional através da modificação genética de mESCs. Os resultados obtidos permitiram verificar que a inibição de Txnip na diferenciação ß-pancreática pode induzir a diferenciação de IPCs com maior expressão de marcadores de células- e mais responsivas ao estímulo de glicose. Além disso, o modelo de zebrafish permitiu elucidar in vivo o papel de Txnip durante a organogênese pancreática, revelando que a inibição desse gene é capaz de aumentar a massa de células-ß através do estimulo de células presentes no ducto extra-pancreático. Dessa forma, a inibição de Txnip pode aprimorar os protocolos para obtenção de IPCs a partir de células-tronco pluripotentes. A exposição crônica a agentes imunossupressores diabetogênicos e a perda de componentes de matriz extracelular durante o isolamento de ilhotas pancreáticas são causas para a perda de funcionalidade do enxerto. Dessa forma, na segunda parte do trabalho, um biomaterial inovador foi desenvolvido, contendo um polímero de laminina (polilaminina, PLn) para o encapsulamento e a imunoproteção de ilhotas pancreáticas. As cápsulas produzidas com o biomaterial desenvolvido, Bioprotect-Pln, são térmica- e mecanicamente estáveis, além de serem biocompatíveis e capazes de imunoproteger ilhotas pancreáticas humanas in vitro. O encapsulamento com Bioprotect-Pln preserva a funcionalidade de ilhotas pancreáticas. Além disso, quando cápsulas vazias de Bioprotect-Pln foram implantadas em camundongos imunocompetentes, houve atenuação da resposta inflamatória ao implante, uma das principais causas para perda de funcionalidade de enxertos encapsulados. Os resultados obtidos indicam que a presença de polilaminina na malha capsular induz uma resposta anti-inflamatória que pode beneficiar a preservação do enxerto de ilhotas pancreáticas encapsuladas. Atualmente, o transplante de ilhotas pancreáticas é visto como a terapia celular mais promissora para atingir a independência de insulina em pacientes de DM1, porém, a aplicabilidade desse transplante ainda é limitada. Este trabalho contribuiu para a elucidação dos mecanismos moleculares que podem aprimorar o processo de diferenciação de célulastronco pluripotentes em IPCs, estabelecendo uma fonte alternativa de células para a terapiade reposição, e, também, estabeleceu um biomaterial inovador, capaz de diminuir a resposta inflamatória ao implante de microcápsulas e de imunoproteger células microencapsuladas. Desta forma, este trabalho contribui para o estabelecimento da terapia de reposição celular para pacientes de DM1
Type 1 diabetes mellitus (DM1) is a disease caused by the autoimmune destruction of insulin-producing pancreatic ß-cells. Pancreatic islet transplantation is a technically simple procedure and an interesting alternative therapy for DM1, however, the limited supply of cadaveric donated pancreas and the need of life-long immunosuppression are factors which limit its applicability. In the present work, two strategies were employed aiming at establishing viable solutions for the factors limiting pancreatic islet transplantation. In the first part of this study, the molecular mechanism which drives differentiation of murine embryonic stem cells (mESCs) into insulin producing cells (IPCs) was analyzed in order to optimize the differentiation process. The Thioredoxin interacting protein (Txnip) gene, which is differentially expressed along -pancreatic differentiation, was selected to undergo a functional analysis by genetically modifying mESCs. The results allowed us to verify that Txnip inhibition during the ß-pancreatic differentiation process can induce differentiation of IPCs displaying higher expression of ß-cell markers and being more responsive to glucose stimuli. In addition, the zebrafish model allowed us to elucidate in vivo the role of Txnip during pancreatic organogenesis, revealing that its inhibition is able to increase the mass of ß-cells through stimulation of extra-pancreatic ductal cells. Therefore, Txnip inhibition may turbinate IPCs differentiation from pluripotent stem cells. The chronic exposure to diabetogenic immunosuppressive agents and the loss of extracellular matrix components during isolation of pancreatic islets are probable causes for the loss of pancreatic islet graft functionality. Therefore, in the second part of this study, an innovative biomaterial was developed by incorporating a laminin polymer (polylaminin, PLn) for the encapsulation and immunoprotection of pancreatic islets. The capsules produced with the novel biomaterial, Bioprotect-Pln, are biocompatible, thermally and mechanically stable and are able to immunoprotect human pancreatic islets in vitro. Encapsulation with Bioprotect-Pln preserves the functionality of pancreatic islets. In addition, when empty Bioprotect-Pln capsules were implanted into immunocompetent mice, an attenuation of the inflammatory response to the implant occurred, this being one of the main causes of encapsulated graft loss. The results indicate that polylaminin addition to the capsular mesh induces an anti-inflammatory response which may favor preservation of the engrafted encapsulated pancreatic islets. Pancreatic islet transplantation is currently seen as the most promising cell therapy to achieve insulin independence in DM1 patients, however, the applicability of this transplant is still limited. This work contributed to the elucidation of the molecular mechanisms which can turbinate the differentiation of pluripotent stem cells into IPCs, establishing an alternative source of cells for the replacement therapy, and, also, established an innovative biomaterial which is able to decrease the inflammatory response to the graft, thereby immunoprotecting the microencapsulated cells. Therefore, this work contributes to the establishment of the cell replacement therapy for DM1 patients