Abstract Background Macrophages have been demonstrated to promote the rupture of the atherosclerotic lesion, a primary cause of ischemic events, by modulating the formation of plaque necrosis. We aimed to search the key factors of genetic macrophages in concerns of carotid ruptured plaques.Methods The database of the gene expressed GSE41571 is retrieved from the Gene Expression Omnibus (GEO) to differentially obtain the measured expression genes (DEGs) by tendering the tool usage of GEO2R. The enriched pathways of ontology at DEGs performances with DAVID interacted the constructions of protein to protein networking by the analysis of STRING and Cytoscape software. And also the identified PPI networks in the significance of hub genes module plugged into respective Cytohubba and MCODE Cytoscape.Result A total of 1481 DEGs in macrophages from ruptured and stable carotid atherosclerotic plaques are noticed, by including 568 up-regulation genes and 913 down-regulation genes. The analyzed GO shows a DEGs are mainly involved in protein ubiquitination, endocytosis, mRNA processing, and mitotic cell cycle. KEGG pathway enrichment analysis revealed that some major protein catabolism pathways, including the ubiquitin-proteasome, autophagy, and endocytosis systems, play key roles in the progressive carotid plaque clotting. The genes of the hub in the PPI network included POLR2E , PPP2R1A , HSP90AA1 , and CBL . The modules were mainly associated with ubiquitin-mediated proteolysis, endocytosis, spliceosome, proteasome.Conclusion Our findings offer novel molecular insights into the mechanisms by which macrophages drive formation and vulnerability of plaques. The candidate DEGs and potential biological pathways might be used as diagnostic targets, and facilitate the development of novel macrophage-targeting therapies for unstable atherosclerosis.
To isolate the collagen phagocytic subpopulation of fibroblast (CPSF) and non-collagen phagocytic subpopulation of fibroblast (nCPSF) and to identify their differentially expressed genes. The CPSF and nCPSF was isolated by using collagen-fluorescein-isothiocynate-latex bead (COL-FITC-LB) phagocytosis technique and FCM sorting method. Microarray analysis was used to screen the differentially expressed genes, which were verified by real-time PCR. CPSF and nCPSF was successfully isolated. Seventeen differentially expressed genes were identified. Compared with nCPSF, the expression of 12 or 5 genes was up-regulated or down-regulated in CPSF. Three of the 12 up-regulated genes were urokinase plasminogen activator receptor-associated protein (uPARAP), cytochrome b-245, beta polypeptide (CYBB) and Hook homolog 1 (HOOK1), which were confirmed by real-time PCR. uPARAP mRNA expression level in CPSF was 2788 times of that in nCPSF. CYBB mRNA expression in CPSF was only 0.85 times of that in nCPSF. HOOK1 mRNA expression in CPSF was 1.96 times of that in nCPSF (P<0.05). A novel method is successfully established to isolate CPSF and nCPSF. uPARAP is the main differentially expressed gene in CPSF and nCPSF, which is obviously involved in the fibroblast collagen phagocytosis. It might be a potential biomarker for treatment of collagen diseases.目的：探讨建立新的分离人成纤维细胞胶原吞噬亚群(CPSF)和成纤维细胞非胶原吞噬亚群(nCPSF)的方法，并筛选两者差异表达基因。方法：根据CPSF和nCPSF吞噬功能的差异，联合应用胶原包被的异硫氰酸荧光乳胶微球(COL-FITC-LB)吞噬技术和流式细胞分离技术分离二者，并通过生物芯片技术和real-time PCR筛选、验证两者差异表达基因，寻找CPSF和nCPSF具有特征性的分子标志物。结果：成功分离CPSF和nCPSF，并对其基因差异性分析，得到17个差异表达基因。与nCPSF比较，CPSF中有12个基因表达上调，5个表达下调。对其中与胶原吞噬功能可能相关的3个基因进行验证，即纤溶酶原激活物受体相关蛋白(urokinase plasminogen activator receptor-associated protein，uPARAP)、细胞色素b-245，β多肽(cytochrome b-245, beta polypeptide，CYBB)和同源物1(Hook homolog 1，HOOK1)，CPSF中uPARAP表达量为nCPSF的2 788倍；CPSF中CYBB表达量为nCPSF的0.85倍；CPSF中HOOK1表达量为nCPSF的1.96倍，差异具有统计学意义(P<0.05)。结论：建立了分离CPSF和nCPSF的新方法。CPSF和nCPSF最主要的差异表达基因是uPARAP，对成纤维细胞胶原吞噬功能具有重要影响，是治疗胶原代谢疾病新的候选靶分子之一。.
Abstract This study investigates aberrant DNA methylations as potential diagnosis and prognosis markers for esophageal squamous-cell carcinoma (ESCC), which if diagnosed at advanced stages has <30% five-year survival rate. Comparing genome-wide methylation sites of 91 ESCC and matched adjacent normal tissues, we identified 35,577 differentially methylated CpG sites (DMCs) and characterized their distribution patterns. Integrating whole-genome DNA and RNA-sequencing data of the same samples, we found multiple dysregulated transcription factors and ESCC-specific genomic correlates of identified DMCs. Using featured DMCs, we developed a 12-marker diagnostic panel with high accuracy in our dataset and the TCGA ESCC dataset, and a 4-marker prognostic panel distinguishing high-risk patients. In-vitro experiments validated the functions of 4 marker host genes. Together these results provide additional evidence for the important roles of aberrant DNA methylations in ESCC development and progression. Our DMC-based diagnostic and prognostic panels have potential values for clinical care of ESCC, laying foundations for developing targeted methylation assays for future non-invasive cancer detection methods.
Objective
To observe the changes in bone mineral density and microstructure parameters in sclerostin (SOST) gene knockout mice treated with glucocorticoid.
Methods
12 4-week-old SOST knockout mice were randomly divided into two groups (n=6): methylprednisolone intervention group [SOM group, methylprednisolone 3 mg/(kg·d), subcutaneous injection], placebo group (SOS group, isovolumetric saline subcutaneous injection). 12 wild-type mice were randomly divided into two groups (n=6): wild-type placebo group (WTS group, isovolumetric saline subcutaneous injection), wild methylprednisolone intervention group [WTM group, methylprednisolone 3 mg/(kg·d), subcutaneous injection]. 12 weeks later, mice were sacrificed and one lumbar vertebra of each mouse was selected for micro-CT analysis.
Results
There was no difference in bone mineral density (BMD), trabecular volume fraction, trabecular number and trabecular thickness between SOM and SOS groups (P>0.05). BMD, trabecular volume fraction, trabecular number and trabecular thickness in SOM and SOS groups were significantly higher than those in WTS and WTM groups (P<0.05). BMD, trabecular volume fraction, trabecular number and trabecular thickness in WTM group were significantly lower than those in WTS group (P<0.05).
Conclusions
Sclerotin gene knockout mice can resist glucocorticoid-induced bone loss and bone microarchitectural deterioeration. The treatment of osteoporosis with SOST/sclerotin as a target will be an effective method in the future.
Key words:
Glucocorticoids; Sclerotin; Gene knockout techniques; Bone density; Mice
Previous data suggest that myostatin has direct effects on the proliferation and differentiation of osteoprogenitor cells. The relationships between serum myostatin, body composition lipids and bone mineral density in postmenopausal women remain unclear. The aim of this study is to elucidate the relationships between serum myostatin, body composition, lipids and bone mineral density in central south Chinese postmenopausal women.A cross-sectional study was conducted in 175 healthy postmenopausal women, aged 51-75 years old. Bone mineral density (BMD) and body composition were measured by double energy X-ray absorptiometry (DXA). Serum myostatin, 25-dihydroxyvitamin D(25OH-D), parathyroid hormone (PTH), bone alkaline phosphatase (BAP) and carboxy-terminal telopeptide of type I collagen (CTX) were measured by enzyme-linked immunoabsorbent assay (ELISA).In contrast to the osteoporotic women, the women without osteoporosis had higher BMI, fat mass and lean mass (P<0.01). The osteoporotic women were older than women without osteoporosis (P<0.01). There were no differences between two groups with regard to serum BAP, CTX, (25OH-D), PTH, lipids and myostatin after adjusted by age. BMD at each site was positively correlated with age at menopause, fat mass and lean mass, and also negatively correlated with age and serum BAP. Serum myostatin was positively correlated with tryglicerides, not correlated with either body composition or BMD at each site.Our data indicated that serum myostatin concentration did not correlate with muscle and bone mass. Further studies are needed to demonstrate the role of myostatin in regulating the bone metabolism.
Cervical cancer is a common malignant tumor in women with increased incidence and younger onset age. As a curable tumor, timely diagnosis and early intervention are critical. Based on the golden standard of cervical tissues pathology examination, we investigated the value of free body of reduced iron protoporphyrin (FH) in uterus epithelial cells for the diagnosis of cervical cancer and precancerous lesions, aiming to provide novel methods for early screening of cervical cancer.A total of 574 women who were screened for cervical cancer according to golden standard of pathology as the reference, were recruited for the analysis of authenticity, reliability, and predictive values of FH. The diagnostic value of FH on cervical carcinoma and precancerous lesion diagnosis was further analyzed.340 individuals had normal cervical or benign lesion by pathology examination, while 155 people had precancerous lesion, among which 79 cases presented early infiltration and infiltrative cancer. In FH screening, 361 and 213 people had negative and positive results, respectively. No significant differences in the results were observed between the two methods in screening cancer and precancerous lesion (p>0.05). FH showed 93.55% sensitivity and 81.94% specificity in diagnosing precancerous lesion, while the sensitivity and specificity for cervical cancer diagnosis were 93.53% and 81.01%, respectively.FH assay was demonstrated to have advantages of high diagnostic value for cervical cancer and precancerous lesion, and might be used for early screening.
To determine the role of Midkine (MDK) in non-invasive detection of bladder cancer (Bca) and the relationship with Ki67.Sixty-five Bca patients and 55 non-Bca patients or healthy volunteers were enrolled and voided urine samples were prospectively obtained on the first day of enrollment. Tissue samples were collected by surgery. MDK and Ki67 expressions were analyzed by immunohistochemistry and Western Blot (WB). Specificity and sensitivity of MDK mRNA testing in the detection of Bca were determined by Receiver Operating Characteristic curve (ROC). The relationship between MDK and Ki67 was also assessed.MDK was overexpressed in Bca tissues than that in the non-cancer tissues. The specificity and sensitivity for MDK mRNA testing in urine in the identification of Bca was 80% and 72.3%. MDK detected 85.7% of high-grade tumors, 87.5% of muscle-invasive tumors and 79.4% of tumors larger than 3 cm in patients without gross hematuria. Microscopic hematuria may even increase the detection rate of Bca by MDK testing. Furthermore, the correlation of MDK and Ki67 was found positive.MDK was overexpressed in Bca tissues and positively correlated with Ki67. MDK might be a potential biomarker for the detection of Bca, especially for those without gross hematuria but with microscopic hematuria.
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