Advancing breast cancer diagnosis with a near-infrared fluorescence imaging smart sensor for estrogen/progesterone receptor detection.

Molecular-genetic imaging has greatly advanced clinical diagnosis and prognosis monitoring. However, the specific visualization of intracellular proteins such as estrogen receptor (ER) and progesterone receptor (PR) remains an elusive goal. Here, we highlight a novel method for selectively detecting ER/PR positive tumors using genetically engineered responsive elements. Our study demonstrates that the double responsive elements of ER/PR exhibit the most sensitivity to the steroid receptors in breast cancers. By utilizing a cationic polymer vector, we constructed a responsive element-fluorescence protein system that can selectively image ER/PR positive breast cancers in murine models under a near-infrared laser. This non-invasive imaging achieved high-resolution detection without death or serious anaphylactic activity in the animals. Our findings suggest that the reporter system consisting of steroid receptor response elements and near-infrared proteins provides a practical system for identifying biomarkers and advancing cancer diagnosis and therapy.

ACAT1-mediated METTL3 acetylation inhibits cell migration and invasion in triple negative breast cancer.

Triple-negative breast cancer (TNBC) is a heterogeneous and aggressive disease with poor prognosis. Acetylation modifications affect a great number of biological processes of malignant tumors. The current study aims at revealing the role of acetylation-related mechanism in TNBC progression. Methyltransferase like-3 (METTL3) was found to be downregulated in TNBC cells via quantitative polymerase chain reaction (qPCR) and western blot analyses. Co-Immunoprecipitation (Co-IP) and GST pulldown assays revealed the interaction between acetyl-CoA acetyltransferase 1 (ACAT1) and METTL3. Through further immunoprecipitation (IP) assay, we determined that ACAT1 stabilizes METTL3 protein via inhibiting the degradation of ubiquitin-proteasome. Functionally, ACAT1 inhibits TNBC cell migration and invasion. Moreover, nuclear receptor subfamily 2 group F member 6 (NR2F6) regulates ACAT1 expression at transcriptional level. Finally, we demonstrated that NR2F6/ACAT/METTL3 axis suppresses the migration and invasion of TNBC cells via METTL3. In conclusion, NR2F6 transcriptionally activates ACAT1 and promotes the suppressive effects of ACAT1-mediated METTL3 acetylation on TNBC cell migration and invasion.

Risk factors of recurrent thyroid nodules after radiofrequency ablation.

Objective: To investigate the risk factors of thyroid nodule recurrence after radiofrequency ablation (RFA). METHODS The medical record information of 120 patients with thyroid nodules admitted to our hospital from June 2019 to April 2022 was retrospectively analysed. All participants received RFA treatment. According to the results of the postoperative thyroid ultrasound examination (USG), the patients were divided into the recurrence group (R, N=16) and the non-recurrence group (NR, N=104). Binary logistic regression analysis was performed to identify the independent risk factors of thyroid nodule recurrence after RFA. The receiver operating characteristic (ROC) curve was used to analyse the value of the forecast of each independent factor and combined model for thyroid nodule recurrence after RFA. Results: During the follow-up period, 16 patients recurred, and the recurrence rate was 13.33%. Univariate regression analysis showed that whether the nodules are solitary (WNS), nodule diameter (ND), the degree of risk of nodular location (DRN), recurrent laryngeal nerve (RLN) injury were associated with thyroid nodule recurrence after RFA (P<0.05). Binary logistic regression analysis showed that WNS, ND, DRN and RLN injury were independent risk factors for the recurrence of thyroid nodules after RFA (P<0.05). ROC analysis of independent factors and combined model showed that solitary nodules, nodule diameter and nodule location risk degree had diagnostic value, while RLN injury had no predictive value. The combined model is more predictive than the independent factors. Conclusions: The risk factors of recurrent thyroid nodules after radiofrequency ablation are related to WNS, ND, DRN and so on, which should be paid attention to and preventive measures should be taken.

Research progress on microRNA-1258 in the development of human cancer.

microRNAs (miRNAs) are small endogenous RNAs composed of 20-22 nucleotides that do not encode proteins, which regulate the expression of downstream genes by targeting the 3' untranslated region of mRNA. Plentiful research has demonstrated that miRNAs participate in the initiation and development of diverse diseases and malignant tumors. miR-1258 exerts great influence on tumors, including tumor growth, distant metastasis, migration, invasion, chemosensitivity, cell glycolysis, apoptosis, and stemness. Interestingly, miR-1258 is a miRNA with explicit functions and has been investigated to act as a tumor suppressor in studies on various types of tumors. With accumulating research on miR-1258, it has been found to be used as a biomarker in the early diagnosis and prognosis prediction of tumor patients. In this review, we outline the development of miR-1258 research, describe its regulatory network, and discuss its roles in cancer. Additionally, we generalize the potential clinical applications of miR-1258. This review offers emerging perspectives and orientations for further comprehending the function of miR-1258 as a diagnostic and prognostic biomarker and potent therapeutic target in cancer.

Construction of the Six-lncRNA Prognosis Signature as a Novel Biomarker in Esophageal Squamous Cell Carcinoma.

Esophageal squamous cell carcinoma (ESCC) is a common malignant gastrointestinal tumor threatening global human health. For patients diagnosed with ESCC, determining the prognosis is a huge challenge. Due to their important role in tumor progression, long non-coding RNAs (lncRNAs) may be putative molecular candidates in the survival prediction of ESCC patients. Here, we obtained three datasets of ESCC lncRNA expression profiles (GSE53624, GSE53622, and GSE53625) from the Gene Expression Omnibus (GEO) database. The method of statistics and machine learning including survival analysis and LASSO regression analysis were applied. We identified a six-lncRNA signature composed of AL445524.1, AC109439.2, LINC01273, AC015922.3, LINC00547, and PSPC1-AS2. Kaplan-Meier and Cox analyses were conducted, and the prognostic ability and predictive independence of the lncRNA signature were found in three ESCC datasets. In the entire set, time-dependent ROC curve analysis showed that the prediction accuracy of the lncRNA signature was remarkably greater than that of TNM stage. ROC and stratified analysis indicated that the combination of six-lncRNA signature with the TNM stage has the highest accuracy in subgrouping ESCC patients. Furthermore, experiments subsequently confirmed that one of the lncRNAs LINC01273 may play an oncogenic role in ESCC. This study suggested the six-lncRNA signature could be a valuable survival predictor for patients with ESCC and have potential to be an auxiliary biomarker of TNM stage to subdivide ESCC patients more accurately, which has important clinical significance.

Corrigendum: Research progress on microRNA-1258 in the development of human cancer.

[This corrects the article DOI: 10.3389/fonc.2022.1024234.].

Identification of Immune-Cell-Related Prognostic Biomarkers of Esophageal Squamous Cell Carcinoma Based on Tumor Microenvironment.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is one of the most fatal cancers in the world. The 5-year survival rate of ESCC is <30%. However, few biomarkers can accurately predict the prognosis of patients with ESCC. We aimed to identify potential survival-associated biomarkers for ESCC to improve its poor prognosis. METHODS: ImmuneAI analysis was first used to access the immune cell abundance of ESCC. Then, ESTIMATE analysis was performed to explore the tumor microenvironment (TME), and differential analysis was used for the selection of immune-related differentially expressed genes (DEGs). Weighted gene coexpression network analysis (WGCNA) was used for selecting the candidate DEGs. Least absolute shrinkage and selection operator (LASSO) Cox regression was used to build the immune-cell-associated prognostic model (ICPM). Kaplan-Meier curve of survival analysis was performed to evaluate the efficacy of the ICPM. RESULTS: Based on the ESTIMATE and ImmuneAI analysis, we obtained 24 immune cells' abundance. Next, we identified six coexpression module that was associated with the abundance. Then, LASSO regression models were constructed by selecting the genes in the module that is most relevant to immune cells. Two test dataset was used to testify the model, and we finally, obtained a seven-genes survival model that performed an excellent prognostic efficacy. CONCLUSION: In the current study, we filtered seven key genes that may be potential prognostic biomarkers of ESCC, and they may be used as new factors to improve the prognosis of cancer.

[Intraoperative Aspiration].

Intraoperative aspiration is a common pulmonary complication in the surgery, anesthesia and position were main factors leading to the operative aspiration. In recent years, perioperative lung protection has attracted wide attention of thoracic surgeons and anesthetist; how to accelerate the process of postoperative rehabilitation, reduce the incidence of related complications and significantly improve the prognosis of patients, these have become a chief goal of surgical treatment. This article will center on operative aspiration and summarize it from anatomy, pathophysiology, manifestation, diagnosis, treatment and prevention.

Weighted Gene Co-expression Network Analysis for RNA-Sequencing Data of the Varicose Veins Transcriptome.

OBJECTIVE: Varicose veins are a common problem worldwide and can cause significant impairments in health-related quality of life, but the etiology and pathogenesis remain not well defined. This study aims to elucidate transcriptomic regulations of varicose veins by detecting differentially expressed genes, pathways and regulator genes. METHODS: We harvested great saphenous veins (GSV) from patients who underwent coronary artery bypass grafting (CABG) and varicose veins from conventional stripping surgery. RNA-Sequencing (RNA-Seq) technique was used to obtain the complete transcriptomic data of both GSVs from CABG patients and varicose veins. Weighted Gene Co-expression network analysis (WGCNA) and further analyses were then carried out with the aim to elucidate transcriptomic regulations of varicose veins by detecting differentially expressed genes, pathways and regulator genes. RESULTS: From January 2015 to December 2016, 7 GSVs from CABG patients and 13 varicose veins were obtained. WGCNA identified 4 modules. In the brown module, gene ontology (GO) analysis showed that the biological processes were focused on response to stimulus, immune response and inflammatory response, etc. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the biological processes were focused on cytokine-cytokine receptor interaction and TNF signaling pathway, etc. In the gray module, GO analysis showed that the biological processes were skeletal myofibril assembly related. The immunohistochemistry staining showed that the expression of ASC, Caspase-1 and NLRP3 were increased in GSVs from CABG patients compared with varicose veins. Histopathological analysis showed that in the varicose veins group, the thickness of vascular wall, tunica intima, tunica media and collagen/smooth muscle ratio were significantly increased, and that the elastic fiber/internal elastic lamina ratio was decreased. CONCLUSION: This study shows that there are clear differences in transcriptomic information between varicose veins and GSVs from CABG patients. Some inflammatory RNAs are down-regulated in varicose veins compared with GSVs from CABG patients. Skeletal myofibril assembly pathway may play a crucial role in the pathogenesis of varicose veins. Characterization of these RNAs may provide new targets for understanding varicose veins diagnosis, progression, and treatment.

Geometry of the Carotid Artery and Its Association With Pathologic Changes in a Chinese Population.

OBJECTIVES: Carotid artery geometry influences blood flow disturbances and is thus an important risk factor for carotid atherosclerosis. Extracellular matrix (ECM) and yes-associated protein (YAP) expression may play essential roles in the pathophysiology of carotid artery stenosis, but the effect of blood flow disturbances of carotid bifurcation location on the ECM is unknown. We hypothesized that carotid artery anatomy and geometry are independently associated with the ECM and YAP expression. METHODS: In this cross-sectional study, 193 patients were divided into two groups: an asymptomatic group (n = 111) and a symptomatic group (n = 82), symptomatic patients presenting with ischemic attack, amaurosis fugax, or minor non-disabling stroke. For all subjects before surgery, carotid bifurcation angle and internal artery angle were measured with computed tomography angiography (CTA), and laminar shear stress was measured with ultrasonography. After surgery, pathology of all plaque specimens was analyzed using hematoxylin and eosin (HE) staining and Movat special staining. Immunohistochemistry was performed to detect expression of YAP in a subset of 30 specimens. RESULTS: Symptomatic patients had increased carotid bifurcation angle and laminar shear stress compared to asymptomatic patients (P < 0.05), although asymptomatic patients had increased internal carotid angle compared to symptomatic patients (P < 0.001). Relative higher bifurcation angles were correlated with increased carotid bifurcation, decreased internal angle, and decreased laminar shear stress. For each change in intervertebral space or one-third of vertebral body height, carotid bifurcation angle changed 4.76°, internal carotid angle changed 6.91°, and laminar shear stress changed 0.57 dynes/cm2. Pathology showed that average fibrous cap thickness and average narrowest fibrous cap thickness were greater in asymptomatic patients than symptomatic patients (P < 0.05). Expression of proteoglycan and YAP protein in symptomatic patients was higher than in asymptomatic patients (P < 0.001), while collagen expression was lower in symptomatic patients than asymptomatic patients (P < 0.05). CONCLUSION: Geometry of the carotid artery and position relative to cervical spine might be associated with ECM and YAP protein expression, which could contribute to carotid artery stenosis.

Exosomes of Endothelial Progenitor Cells Inhibit Neointima Formation After Carotid Artery Injury.

BACKGROUND: Exosomes released from endothelial progenitor cells (EPCs) play a protective role in various disease models. Both endothelial cell (EC) damage and smooth muscle cell (SMC) proliferation are involved in the pathological process of restenosis after angioplasty and stenting. Few studies have focused on the therapeutic role of exosomes in EC damage and SMC proliferation. In this study, we sought to investigate the effect of exosomes released by human fetal aorta-derived EPCs on the rat carotid artery balloon injury model in vivo. We also sought to determine the effect of exosomes on both ECs and SMCs in vitro. METHODS: Exosomes (Exo group) or saline (Con group) were injected in rat carotid balloon injury model animals. The rats were sacrificed after 2, 4, 14, and 28 d, and injured carotid specimens were collected for Evans blue staining, hematoxylin-eosin staining, and immunohistochemistry. RESULTS: When the Con group and the Exo group were compared, the reendothelialized areas were not significantly different after 2 or 4 d, as shown by Evans blue staining. The hematoxylin-eosin results showed that the intimal to medial area ratio was slightly but not significantly higher in the Exo group after 2 and 4 d. The immunohistochemistry results showed that the proliferation of SMCs was slightly higher in the Exo group after 2 and 4 d, but the difference was not significant. The reendothelialization area of the Con group was significantly smaller than that of the Exo group at day 14. Both the intimal to medial area ratio and SMC proliferation in the Exo group were significantly smaller than those of the Con group at 14 or 28 d. In the in vitro study, exosome treatment significantly enhanced the proliferation and migration of both ECs and SMCs. CONCLUSIONS: Exosomes derived from EPCs could inhibit neointimal hyperplasia after carotid artery injury in rats. The protective effect of exosomes may manifest through the promotion of EC repair rather than direct suppression of proliferation and migration of smooth muscles cells.

Programmed Cell Death Protein-1 Predicts the Recurrence of Breast Cancer in Patients Subjected to Radiotherapy After Breast-Preserving Surgery.

Radiotherapy is the most important component of the comprehensive treatment of breast cancer, and immunocompromised patients respond with lower response rate. However, the role of programmed cell death protein-1, a critical immune molecule, in recurrence of breast cancer subjected to radiotherapy is unknown. A retrospective analysis was designed to explore the relevance. A number of 42 patients with early-stage breast cancer undergoing breast-conserving surgery and postoperative radiotherapy (18 recurrence and 24 nonrecurrence) were recruited, and clinical data were obtained. Immunohistochemistry was employed to detect programmed cell death protein-1, and Kaplan-Meier curves were used to analyze recurrence-free survival. The expression of programmed cell death protein-1 was higher in the recurrence group than recurrence-free group ( P < .05). Meanwhile, the recurrence-free mean survival was significantly longer in programmed cell death protein-1 low-expression group (68 months) than that in programmed cell death protein-1 high-expression group (56 months). In addition, the levels of T lymphocytes were obviously lower in patients with breast cancer than healthy group, and natural killer showed an opposite tendency. CD4+ decreased significantly after 1 week radiotherapy and recovered rapidly 3 weeks after radiotherapy. Compared to recurrence-free group, the increment of T lymphocytes were inadequate in recurrence group. These experimental results indicated that the expression of programmed cell death protein-1 in tumor-infiltrating lymphocytes is related to immune disorder and recurrence of patients undergoing breast-preserving surgery and radiotherapy.

Is Human-induced Pluripotent Stem Cell the Best Optimal?

OBJECTIVE: Since the advent of induced pluripotent stem cell (iPSC) technology a decade ago, enormous progress has been made in stem cell biology and regenerative medicine. Human iPSCs have been widely used for disease modeling, drug discovery, and cell therapy development. In this review, we discuss the progress in applications of iPSC technology that are particularly relevant to drug discovery and regenerative medicine, and consider the remaining challenges and the emerging opportunities in the field. DATA SOURCES: Articles in this review were searched from PubMed database from January 2014 to December 2017. STUDY SELECTION: Original articles about iPSCs and cardiovascular diseases were included and analyzed. RESULTS: iPSC holds great promises for human disease modeling, drug discovery, and stem cell-based therapy, and this potential is only beginning to be realized. However, several important issues remain to be addressed. CONCLUSIONS: The recent availability of human cardiomyocytes derived from iPSCs opens new opportunities to build in vitro models of cardiac disease, screening for new drugs and patient-specific cardiac therapy.

Surgical treatment of recurrent varicose veins in the lower limbs associated with endovascular treatment of iliac vein stenosis.

OBJECTIVE: We present our experience with endovascular surgery for recurrent varicose veins (RVV) of the lower limbs combined with the iliac vein compression syndrome (IVCS). MATERIALS AND METHODS: This study was a retrospective analysis of 6 patients with RVVs combined with IVCS who were admitted to our hospital between January 2007 and December 2014. Transfemoral venography was performed to confirm IVCS. Balloon dilation and stent placement were successful in all 6 patients. The varicose veins were treated by traditional surgery after the endovascular therapy. The visual analog pain scale (VAS) score and venous clinical severity score (VCSS) were collected before surgery and at 6-months follow-up, and were analyzed using the paired student t-test. Patency of the iliac vein was assessed via duplex Doppler ultrasound. RESULTS: The rate of technical success was 100%. There was a significant (p < .001) improvement in VCSS postoperatively. During the 6-month follow-up period, no RVVs were observed and the rate of iliac vein patency was 100%. Importantly, VAS ratings also decreased significantly (p < .001) during the follow-up. CONCLUSION: Endovascular surgery for IVCS combined with traditional surgery focused on varicose veins is an effective procedure for treating RVVs of the lower limbs associated with IVCS within 6 months.

Long-term Outcomes of Paclitaxel-Eluting Versus Sirolimus-Eluting Stent for Percutaneous Coronary Intervention: AMeta-Analysis.

The relative long-term efficacy and safety of sirolimus-eluting stents (SES) compared with paclitaxel-eluting stents (PES) in multiple comparative studies remains controversial. This report evaluates 29 randomized trials with 18,379 patients in whom long-term (more than 1 year) outcomes were evaluated. The primary outcomes were target lesion revascularization (TLR) and the secondary end points were death, cardiac death, myocardial infarction (MI), major adverse cardiac events (MACEs), target vessel revascularization (TVR) and stent thrombosis (ST). In comparison with PES, SES significantly reduced the long-term risk of TLR (RR=0.68; 95% CI=0.57 to 0.80, p<0.001), TVR (RR=0.69; 95% CI= 0.60 to 0.79, p<0.001) and MACE (RR=0.82; 95% CI= 0.77 to 0.88, p<0.001), while there were no significant difference with respect to death, cardiac death, MI and STbetween the two groups. SES performance was significantly better for reducing the former three outcomes and comparable for the majority of the secondary end points when compared against PES.

Diagnostic Accuracy Study of Intraoperative and Perioperative Serum Intact PTH Level for Successful Parathyroidectomy in 501 Secondary Hyperparathyroidism Patients.

Parathyroidectomy (PTX) is an effective treatment for severe secondary hyperparathyroidism (SHPT); however, persistent SHPT may occur because of supernumerary and ectopic parathyroids. Here a diagnostic accuracy study of intraoperative and perioperative serum intact parathyroid hormone (iPTH) was performed to predict successful surgery in 501 patients, who received total PTX + autotransplantation without thymectomy. Serum iPTH values before incision (io-iPTH0), 10 and 20 min after removing the last parathyroid (io-iPTH10, io-iPTH20), and the first and fourth day after PTX (D1-iPTH, D4-iPTH) were recoded. Patients whose serum iPTH was >50 pg/mL at the first postoperative week were followed up within six months. Successful PTX was defined if iPTH was <300 pg/mL, on the contrary, persistent SHPT was regarded. There were 86.4% patients underwent successful PTX, 9.8% remained as persistent SHPT and 3.8% were undetermined. Intraoperative serum iPTH demonstrated no significant differences in two subgroups with or without chronic hepatitis. Receiver operating characteristic (ROC) curves showed that >88.9% of io-iPTH20% could predict successful PTX (area under the curve [AUC] 0.909, sensitivity 78.6%, specificity 88.5%), thereby avoiding unnecessary exploration to reduce operative complications. D4-iPTH >147.4 pg/mL could predict persistent SHPT (AUC 0.998, sensitivity 100%, specificity 99.5%), so that medical intervention or reoperation start timely.

Parathyroidectomy and heart rate variability in patients with stage 5 CKD.

BACKGROUND AND OBJECTIVES: Lower heart rate variability implies increased risk of cardiovascular disease. This study aimed to evaluate the relationship between mineral metabolism and heart rate variability and longitudinal changes of heart rate variability after parathyroidectomy in stage 5 CKD patients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This cross-sectional study included 118 stage 5 CKD patients, 87 controls, and a prospective study in two subgroups classified as successful (n=17) and unsuccessful (n=4) parathyroidectomy follow-up enrolled from March of 2011 to December of 2012. Blood examination and 24-hour Holter for heart rate variability were measured. RESULTS: Most heart rate variability indices were lower in stage 5 CKD patients. In multivariate stepwise regression models, serum intact parathyroid hormone was correlated with mean normal-to-normal R-R intervals, mean heart rate, and very low frequency, serum calcium was correlated with SD of 5-minute average of normal R-R intervals, and serum phosphorus was correlated with very low frequency and low frequency/high frequency. Compared with baseline, the successful parathyroidectomy subgroup had significant improvements in mean normal-to-normal R-R intervals, mean heart rate, SD of normal-to-normal R-R intervals, SD of 5-minute average of normal R-R intervals, very low frequency, high frequency, and low frequency/high frequency. There was no significant change of heart rate variability in patients after unsuccessful parathyroidectomy. CONCLUSIONS: Disorders of mineral metabolism are associated with decreased heart rate variability in stage 5 CKD. Successful parathyroidectomy may contribute to reverse this cardiovascular disease risk in severe secondary hyperparathyroidism patients.

SOD mimetic improves the function, growth, and survival of small-size liver grafts after transplantation in rats.

BACKGROUND: Small-for-size syndrome (SFSS) may occur when graft volume is less than 45% of the standard liver volume, and it manifests as retarded growth and failure of the grafts and more mortality. However, its pathogenesis is poorly understood, and few effective interventions have been attempted. AIMS: The present study aimed to delineate the critical role of oxidant stress in SFSS and protective effects of a superoxide dismutase mimetic, Mn(III)tetrakis(4-benzoic acid)porphyrin chloride (MnTBAP), on graft function, growth, and survival in the recipient rats. METHODS: Small size graft liver transplantation (SSGLT) was performed to determine the survival, graft injury, and growth. MnTBAP was administered in SSGLT recipients (SSGLT+MnTBAP). RESULTS: Serum alanine aminotransferase levels were sustained higher in SSGLT recipients, which were correlated with an increased apoptotic cell count and hepatocellular necrosis in liver sections. Malondialdehyde content, gene expression of tumor necrosis factor α and interleukin 1ß, and DNA binding activity of nuclear factor-κB in the grafts were increased significantly in SSGLT recipients compared with sham-operated controls. Both phosphorylated p38 mitogen-activated protein kinase and nuclear c-Jun were increased in SSGLT. All these changes were strikingly reversed by the administration of MnTBAP, with an increase in serum superoxide dismutase activity. Moreover, in situ bromodeoxyuridine incorporation demonstrated that graft regeneration was much more profound in the SSGLT+MnTBAP group than in the SSGLT group. Finally, the survival of recipients with MnTBAP treatments was significantly improved. CONCLUSIONS: Enhanced oxidant stress with activation of the p38/c-Jun/nuclear factor-κB signaling pathway contributes to SFSS-associated graft failure, retarded graft growth, and poor survival. MnTBAP effectively reversed the pathologic changes in SFSS-associated graft failure.

miRNA regulation of liver growth after 50% partial hepatectomy and small size grafts in rats.

BACKGROUND: The molecular mechanisms underlying the growth of small size grafts and the remaining livers are poorly understood. MicroRNAs (miRNAs) negatively modulate expression of genes that are involved in cellular function and metabolism. The aim of this study is to identify critical miRNA species that modulate the growth of small grafts and the remaining livers after partial hepatectomy (PH). METHODS: Small size graft liver transplantation was performed in rats. Liver tissue was harvested after transplant or PH for the determination of miRNA expression profile, and the data were confirmed by quantitative reverse-transcriptase polymerase chain reaction. The genes involved in cell cycle and proliferation were analyzed by quantitative reverse-transcriptase polymerase chain reaction and immunohistochemical staining. RESULTS: Compared with control liver, miR_122a, Let_7b, and miR_26a were reduced by more than 90% in 45% volume grafts. In the remaining livers after 50% PH, 30 miRNAs were down-regulated by more than 50%, and among them, miR_22a, miR_26a, miR_30b, Let_7f, and Let_7g were markedly decreased. A negative correlation existed between down-regulated miRNAs and highly up-regulated genes involved in cell cycle and proliferation in the remaining livers. Moreover, overexpression of miR_26a markedly down-regulated cyclin E2 protein levels and significantly decreased proliferation of HepG2 cells. CONCLUSION: Down-regulated miRNAs play a pivotal role in promoting the growth of small size grafts and the remaining livers. The negative correlation between down-regulated miRNAs and up-regulated genes suggests that these specific miRNAs participate in the modulation of a growth response in both living donors and small size graft recipients.

Immunohistochemical staining of cancer stem cell markers in hepatocellular carcinoma.

BACKGROUND: Cancer stem cells (CSCs) are thought to be a critical subpopulation in tumor development, progression, metastasis and recurrence, and the identification of these cells is an initial step in understanding their role in oncogenesis and in seeking valuable markers for diagnosis or development of targeting therapeutics. AIMS: To identify CSCs in hepatocellular carcinoma (HCC) specimens and define their tissue specificity. METHODS: Immunohistochemical staining of CSC markers: CD44, CD90, CD133 and aldehyde dehydrogenase (ALDH) was performed in 25 HCC specimens, 4 hepatoblastomas, 8 peri-malignant tissues, and 19 cases of viral hepatitis. RESULTS: The positivity of CD44 staining in HCC specimens was significantly lower than in viral hepatitis specimens. The positive rate of CD133 in HCC was similar to viral hepatitis specimens. CD133(+) cells were largely localized to ALDH-positive cells in HCC as revealed by confocal microscopy. In contrast, the co-expression of both markers was visualized within vessels or in the portal areas in viral hepatitis. Moreover, among 7 liver specimens adjacent to HCC tissue, 3-6 samples were positive for CD44, CD90, CD133 and ALDH, especially in dysplastic cells. One of 4 hepatoblastoma cases was positive for all these markers; whereas, the other three specimens were negative for all these CSC markers. CONCLUSIONS: In HCC and dysplastic tissues, clusters of CD133(+)/ALDH(high) cells were identified. The use of cancer stem cell markers to screen tissues with chronic liver diseases provides limited guidance in the identification of malignant cells.