Patterns, Risk Factors, and Outcomes of Recurrence After Hepatectomy for Hepatocellular Carcinoma with and without Microvascular Invasion.

Purpose: The patterns and risk factors of postsurgical recurrence of patient with hepatocellular carcinoma (HCC) with microvascular invasion (MVI) are not clarified. This study aimed to decipher and compare the postoperative recurrent patterns and the risk factors contributing to recurrence between MVI positive (MVI(+)) and MVI negative (MVI(-)) HCC after hepatectomy. Patients and methods: Patients with HCC who underwent hepatectomy in three Chinese academic hospitals between January 1, 2009, and December 31, 2018, were enrolled. Recurrent patterns included early (≤2 years) or late (>2 years) recurrence, recurrent sites and number, and risk factors of recurrence were compared between the MVI(+)and MVI(-) groups by propensity score-matching (PSM). Results: Of 1756 patients included, 581 (33.1%) were MVI(+), and 875 (49.8%) patients developed early recurrence. Compared with the MVI(-) group, the MVI(+) group had a higher 2-year recurrence rate in the PSM cohort (hazard ratio [HR], 1.82; 95% confidence interval [CI], 1.59-2.10; P < 0.001), and more patients with multiple tumor recurrence. Patients with early recurrence in the MVI(+) group had a worse overall survival (OS) than those in the MVI(-) group (HR, 1.24; 95% CI, 1.02-1.50; P = 0.034). Resection margin (RM) ≤1.0 cm is a surgical predictor of early recurrence for the MVI(+) group (HR, 0.68; 95% CI, 0.54-0.87; P = 0.002), but not for the MVI(-) group. Conclusion: Compared to MVI(-) HCC, MVI(+) HCC tends to be early, multiple recurrence and lung and lymph node metastasis after resection. RM ≤1.0 cm is a surgical risk factor of early recurrence for patient with MVI.

Small extracellular vesicles derived from human mesenchymal stem cells prevent Th17-dominant neutrophilic airway inflammation via immunoregulation on Th17 cells.

Type 17 helper T cells (Th17)-dominant neutrophilic airway inflammation is critical in the pathogenesis of steroid-resistant airway inflammation such as severe asthma. Small extracellular vesicles (sEV) derived from human mesenchymal stem cells (MSCs) display extensive therapeutic effects and advantages in many diseases. However, the role of MSC-sEV in Th17-dominant neutrophilic airway inflammation and the related mechanisms are still poorly studied. Here we found that MSC-sEV significantly alleviated the infiltration of inflammatory cells in peribronchial interstitial tissues and reduced levels of inflammatory cells, especially neutrophils, in bronchoalveolar lavage fluids (BALF) of mice with neutrophilic airway inflammation. Consistently, MSC-sEV significantly decreased levels of IL-17A in BALF and Th17 in lung tissues. Furthermore, we found that labelled MSC-sEV were taken up by human CD4+ T cells most obviously at 12 h after incubation, and distributed mostly in mouse lungs. More importantly, potential signaling pathways involved in the MSC-sEV mediated inhibition of Th17 polarization were found using RNA sequencing. Using Western blot, JAK2-STAT3 pathway was identified as an important role in the inhibition of Th17 polarization by MSC-sEV. We found that proteins in MSC-sEV were mostly involved in the therapeutic effects of MSC-sEV. In total, our study suggested that MSC-sEV could be a potential therapeutic strategy for the treatment of neutrophilic airway inflammation.

Mesenchymal stromal cells-derived small extracellular vesicles modulate DC function to suppress Th2 responses via IL-10 in patients with allergic rhinitis.

Mesenchymal stromal cells (MSCs) are well known for their immunoregulatory roles on allergic inflammation particularly by acting on T cells, B cells, and dendritic cells (DCs). MSC-derived small extracellular vesicles (MSC-sEV) are increasingly considered as one of the main factors for the effects of MSCs on immune responses. However, the effects of MSC-sEV on DCs in allergic diseases remain unclear. MSC-sEV were prepared from the induced pluripotent stem cells (iPSC)-MSCs by anion-exchange chromatography, and were characterized with the size, morphology, and specific markers. Human monocyte-derived DCs were generated and cultured in the presence of MSC-sEV to differentiate the so-called sEV-immature DCs (sEV-iDCs) and sEV-mature DCs (sEV-mDCs), respectively. The phenotypes and the phagocytic ability of sEV-iDCs were analyzed by flow cytometry. sEV-mDCs were co-cultured with isolated CD4+ T cells or peripheral blood mononuclear cells (PBMCs) from patients with allergic rhinitis. The levels of Th1 and Th2 cytokines produced by T cells were examined by ELISA and intracellular flow staining. And the following mechanisms were further investigated. We demonstrated that MSC-sEV inhibited the differentiation of human monocytes to iDCs with downregulation of the expression of CD40, CD80, CD86, and HLA-DR, but had no effects on mDCs with these markers. However, MSC-sEV treatment enhanced the phagocytic ability of mDCs. More importantly, using anti-IL-10 monoclonal antibody or IL-10Rα blocking antibody, we identified that sEV-mDCs suppressed the Th2 immune response by reducing the production of IL-4, IL-9, and IL-13 via IL-10. Furthermore, sEV-mDCs increased the level of Treg cells. Our study identified that mDCs treated with MSC-sEV inhibited the Th2 responses, providing novel evidence of the potential cell-free therapy acting on DCs in allergic airway diseases.

Plasma EVs Display Antigen-Presenting Characteristics in Patients With Allergic Rhinitis and Promote Differentiation of Th2 Cells.

Background: Allergic rhinitis (AR) is characterized by IgE-mediated mucosa response after exposure to allergens. Extracellular vesicles (EVs) are nano-size vesicles containing biological cargos for intercellular communications. However, the role of plasma EVs in pathogenesis of AR remains largely unknown. Methods: Plasma EVs from patients with AR were isolated, quantified, and characterized. The expression of Der p 1 and antigen-presenting molecules on EVs was determined by Western blot, flow cytometry, or ELISA. PKH26- and CFSE (carboxyfluorescein succinimidyl ester)-stained AR-EVs were used to determine the uptake of EVs by CD4+T cells and their effects on CD4+T cell proliferation, respectively. Results: Plasma EVs in healthy control (HC) and AR patients were similar in the concentration of particles, expression for specific EV markers, and both had structural lipid bilayer. However, the levels of Der p 1 on plasma EVs from both mild and moderate-severe AR patients were significantly higher than that on HC. The levels of antigen-presenting molecules on plasma EVs were similar from three subjects. Moreover, levels of Der p 1 on EVs in plasma, but not nasal secretion, were significantly associated with the symptom score of AR patients and level of plasma IL-13. Additionally, plasma EVs from patients with AR promoted the development of Th2 cells, while no effect was found on CD4+ T-cell proliferation. Conclusions: Plasma EVs derived from patients with AR exhibited antigen-presenting characteristics and promoted differentiation of Th2 cells, thus providing novel understanding of the pathogenesis of AR.

Mesenchymal stem cells regulate type 2 innate lymphoid cells via regulatory T cells through ICOS-ICOSL interaction.

Group 2 innate lymphoid cells (ILC2s) are recognized as key controllers and effectors of type 2 inflammation. Mesenchymal stem cells (MSCs) have been shown to alleviate type 2 inflammation by modulating T lymphocyte subsets and decreasing TH 2 cytokine levels. However, the effects of MSCs on ILC2s have not been investigated. In this study, we investigated the potential immunomodulatory effects of MSCs on ILC2s in peripheral blood mononuclear cells (PBMCs) from allergic rhinitis patients and healthy subjects. We further investigated the mechanisms involved in the MSC modulation using isolated lineage negative (Lin- ) cells. PBMCs and Lin- cells were cocultured with induced pluripotent stem cell-derived MSCs (iPSC-MSCs) under the stimulation of epithelial cytokines IL-25 and IL-33. And the ILC2 levels and functions were examined and the possible mechanisms were investigated based on regulatory T (Treg) cells and ICOS-ICOSL pathway. iPSC-MSCs successfully decreased the high levels of IL-13, IL-9, and IL-5 in PBMCs in response to IL-25, IL-33, and the high percentages of IL-13+ ILC2s and IL-9+ ILC2s in response to epithelial cytokines were significantly reversed after the treatment of iPSC-MSCs. However, iPSC-MSCs were found directly to enhance ILC2 levels and functions via ICOS-ICOSL interaction in Lin- cells and pure ILC2s. iPSC-MSCs exerted their inhibitory effects on ILC2s via activating Treg cells through ICOS-ICOSL interaction. The MSC-induced Treg cells then suppressed ILC2s by secreting IL-10 in the coculture system. This study revealed that human MSCs suppressed ILC2s via Treg cells through ICOS-ICOSL interaction, which provides further insight to regulate ILC2s in inflammatory disorders.

Long non-coding RNA GAS5 antagonizes the chemoresistance of pancreatic cancer cells through down-regulation of miR-181c-5p.

OBJECTIVE: To explore the core mechanism of long non-coding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) in the regulation of multidrug resistance of pancreatic cancer cells. METHODS: mRNA levels of GAS5, miR-181c-5p and Hippo pathway related genes were detected by quantitative real-time PCR (qRT-PCR). Protein levels of MDR-1, MST1, YAP and TAZ were measured by western blot. Cell viability was detected by MTT assay. The combination between GAS5 and miR-181c-5p was confirmed by RNA pull-down and RNA immunoprecipitation (RIP) assay. We also established pancreatic cancer-bearing mice model and analyzed tumor volumes. RESULTS: Our data showed GAS5 expression was significantly down-regulated, miR-181c-5p expression was significantly up-regulated in pancreatic cancer cells. Besides, Overexpresson of GAS5 obviously inhibited cell viability, while GAS5 knockdown showed the opposite outcome. Additionally, we also found that GAS5 negatively regulated miR-181c-5p, and miR-181c-5p dramatically promoted pancreatic cancer cell chemoresistance through inactivating the Hippo signaling. GAS5 regulated chemoresistance and Hippo pathway of pancreatic cancer cells via miR-181c-5p/Hippo. Finally, we confirmed that overexpression of GAS5 inhibited tumor growth in pancreatic cancer-bearing mice model. CONCLUSION: GAS5 regualtes Hippo signaling pathway via miR-181c-5p to antagonize the development of multidrug resistance in pancreatic cancer cells.

Long non-coding RNA GAS5 suppresses pancreatic cancer metastasis through modulating miR-32-5p/PTEN axis.

BACKGROUND: Long non-coding RNA growth arrest-specific transcript 5 (lncRNA GAS5) is a well-known tumor suppressor in the pathogenesis of a variety of human cancers. The precise role of GAS5 in pancreatic cancer (PC) progression is currently unknown, so the aim of this study was to explore the functional participation of GAS5 in PC metastasis. METHODS: The expression changes of GAS5, miR-32-5p and PTEN in human PC specimens and cell lines were compared by means of molecular biology methods. Transfection of the recombinant plasmid was applied to modulate the expression levels of the target genes. RIP and RNA pull-down assays were designed to investigate the interaction between GAS5 and miR-32-5p. The effect of GAS5 and miR-32-5p on PC progression was assessed with cell proliferation, migration, invasion and apoptosis in vitro. RESULTS: GAS5 and PTEN protein were decreased in human PC tissues and cells, but miR-32-5p was increased. GAS5 induction greatly inhibited the proliferation, migration and invasion of PC cells PANC-1 and BxPC-3 in vitro and simultaneously induced cell apoptosis. Moreover, GAS5 positively regulated the expression of PTEN through miR-32-5p. Furthermore, GAS5 suppressed the proliferation, migration and invasion of PC cells through regulating miR-32-5p/PTEN axis. Additionally, this finding was further supported by the results of in vivo experiments. CONCLUSION: GAS5 could positively regulate PTEN-induced tumor-suppressor pathway via miR-32-5p, thereby suppressing PC metastasis.

Primary nasopharyngeal polyps: a case series on a rare clinical entity.

Primary non-neoplastic polyps originating from the nasopharynx have not been reported in the English language literature. We present the clinical and histopathological features of three primary nasopharyngeal polyps. Clinical data of three patients with primary nasopharyngeal polyps treated at the Department of Otolaryngology, The First Affiliated Hospital of Sun Yat-sen University between 2005 and 2015 were analyzed and presented. Three male patients from 45 to 63 years presented with nasopharyngeal masses. CT or MRI examination showed nasopharyngeal space-occupying lesions. Two patients were initially diagnosed with nasopharyngeal angiofibroma and one patient with nasopharyngeal carcinoma. After surgical excision, based on the histological examination, the tissue masses were all diagnosed as inflammatory polyps. Histologically, the polyps demonstrated significant oedema, collagen deposition, leukocytic infiltration, and epithelial remodelling. Primary nasopharyngeal polyps represent a distinct clinical entity and should be considered in the differential diagnosis of nasopharyngeal masses.

Differential Expression Patterns of EGF, EGFR, and ERBB4 in Nasal Polyp Epithelium.

Epidermal growth factor receptors play an important role in airway epithelial cell growth and differentiation. The current study investigates the expression profiles of EGF, EGFR and ERBB4 in patients with nasal polyps (NP), and their response to glucocorticosteroid (GC) treatment. Fifty patients with NP (40 without GC treatment and 10 with oral GC) and 20 control subjects with septal deviation were recruited into the study. Protein levels of EGF, EGFR, and ERBB4 were evaluated by immune-staining. In healthy nasal epithelium, EGF and EGFR localized within p63+ basal cells, while ERBB4 localized within ciliated cells. GC-naïve NP epithelium showed weak expression of EGF in 90% of samples versus 5% of controls. EGFR was significantly increased in the epithelium with basal cell hyperplasia from GC-naïve NPs (78%, 31/40) compared to controls (23%, 4/17). EGFR was also found in some degranulating goblet cells. ERBB4 expression was significantly higher in hyperplastic epithelium from GC-naïve NPs (65%, 26/40) than in controls (6%, 1/17). GC treatment restored the EGF expression and normalized the EGFR and ERBB4 expression in NPs. Differential expression patterns of EGF, EGFR, and ERBB4 are essential in epithelial restitution and remodeling in nasal epithelium.

Immune responses to different patterns of exposure to ovalbumin in a mouse model of allergic rhinitis.

Allergic rhinitis (AR) has been a significant healthcare burden on individuals and society. However, the detailed effect of different patterns of allergen exposure on the development of AR remains controversial. A mouse model of AR was established to address the complex relationships between allergen exposure and the development of AR. Allergic symptom, OVA-specific IgE in serum and nasal lavage fluid, allergic inflammation in nasal tissues were evaluated after intranasal sensitization and challenge of ovalbumin (OVA) in mice treated with two different doses of allergen for different sensitized durations. Exposure to different doses and sensitized durations of OVA were capable of inducing allergic nasal response. Repetitive OVA exposure in the sensitization phase induced the recruitment of eosinophils and goblet cell hyperplasia. The level of OVA-specific IgE in serum depended on OVA exposure and was mediated in a duration-related manner. In addition, mice treated with low-dose OVA for prolonged duration manifested the major features of human local allergic rhinitis. There were dose- and duration-related effects of allergen exposure on the development of AR. LAR was associated with repetitive exposure to low-dose allergen. Thus, allergen avoidance should be an important aim of AR management.

Epidemiological characterization and risk factors of allergic rhinitis in the general population in Guangzhou City in china.

The prevalence of allergic rhinitis (AR) in China has increased with an apparent geographic variation. The current study aims to investigate the AR prevalence/classification, diagnosis/treatment conditions, trigger factors, and risk factors in the general population of Guangzhou, the third biggest city in China. A cross-sectional survey was performed in the citizens in Guangzhou from December 2009 to March 2010 by using a stratified multistage cluster sampling method. All subjects were asked to complete a comprehensive questionnaire via a face to face interview. A total of 9,899 questionnaires were valid. The prevalence rate of AR in the general population of Guangzhou was 6.24%, with a significant higher prevalence in urban area (8.32%) versus rural area (3.43%). Among the AR subjects, most (87%) were diagnosed with intermittent AR and 87% suffered from moderate-severe symptoms. High percentages of the AR patients did not have previously physician-based diagnosis (34%) or specific medical treatment (55%). Morning time, winter season, and cold air were the most common trigger factors of AR. Family history of AR, current living place, living place during babyhood, smoking, home renovation, and pet ownership were the significant risk factors associated with AR prevalence in the population. The study demonstrated comprehensive epidemiological and clinical information about the AR in Guangzhou population. Change of living environment and lifestyles had strong impacts on the prevalence of AR. Public health policies should help the patients benefit from a proper diagnosis/treatment and specifically target the local risk factors, in order to control the AR incidence.

Removal of tracheobronchial foreign bodies via suspension laryngoscope and Hopkins telescope in infants.

OBJECTIVES: Tracheobronchial foreign body aspiration is a life-threatening accident in infants, and is still a formidable clinical emergency to both otorhinolaryngologists and anesthesiologists. In this study, we attempted to assess the safety and ease of tracheobronchial foreign body removal in infants via suspension laryngoscopy and Hopkins telescopy under general anesthesia with endotracheal intubation. METHODS: The retrospective clinical study from 2006 to 2010 included 50 infants with foreign body aspiration, of whom 35 underwent suspension laryngoscopy and Hopkins telescopy and the other 15 underwent rigid bronchoscopy. All of the procedures were under general anesthesia with endotracheal intubation. RESULTS: All of the patients underwent temporary extubation. The foreign body was successfully removed in 46 cases and was not found in the other 4 cases. The mean operation time in the rigid bronchoscopy group was 13.20+/-9.01 minutes, and that in the Hopkins telescopy group was 5.79+/-3.54 minutes. The oxygen saturation level was below 90% in 17 cases, of which 7 were in the rigid bronchoscopy group and 10 were in the Hopkins telescopy group. The vital signs, including the partial pressure of carbon dioxide in expiratory gas and the heart rate, were stable in all cases. CONCLUSIONS: Foreign body removal in infants via suspension laryngoscopy and Hopkins telescopy under general anesthesia with endotracheal intubation should be promoted, since it is relatively safe and easy for both anesthesiologists and otorhinolaryngologists to perform and has a remarkable success rate.

Suppressed fuzzy-soft learning vector quantization for MRI segmentation.

OBJECTIVE: A self-organizing map (SOM) is a competitive artificial neural network with unsupervised learning. To increase the SOM learning effect, a fuzzy-soft learning vector quantization (FSLVQ) algorithm has been proposed in the literature, using fuzzy functions to approximate lateral neural interaction of the SOM. However, the computational performance of FSLVQ is still not good enough, especially for large data sets. In this paper, we propose a suppressed FSLVQ (S-FSLVQ) using suppression with a parameter learning schema. We then apply the S-FSLVQ to MRI segmentation and compare it with several existing methods. METHODS AND MATERIALS: The proposed S-FSLVQ algorithm and some existing methods, such as FSLVQ, generalized LVQ, revised generalized LVQ and alternative LVQ, are compared using numerical data and MRI images. The numerical data are generated by a mixture of normal distributions. The MRI data sets are from a 2-year-old female patient who was diagnosed with retinoblastoma of her left eye, a congenital malignant neoplasm of the retina with frequent metastasis beyond the lacrimal cribrosa. To evaluate the performance of these algorithms, two criteria for accuracy and computational efficiency are used. RESULTS: Comparing S-FSLVQ with FSLVQ, generalized LVQ, revised generalized LVQ and alternative LVQ, the numerical results indicate that the S-FSLVQ algorithm is better than the other algorithms in accuracy and computational efficiency. Moreover, the proposed S-FSLVQ can reduce the computation time and increase accuracy compared to existing methods in segmenting these ophthalmological MRIs. CONCLUSIONS: The proposed S-FSLVQ is a good competitive learning algorithm that is very suitable for segmenting the ophthalmological MRI data sets. Therefore, the S-FSLVQ algorithm is highly recommended for use in MRI segmentation as an aid for supportive diagnoses.

[Relationship between allergic symptoms and RORC2 and IL-17 in patients with allergic rhinitis].

OBJECTIVE: To evaluate the relationship between allergic symptoms and retinoic acid-related orphan receptor variant 2 (RORC2) and interleukin (IL) 17 in patients with allergic rhinitis (AR). METHODS: Blood sample, nasal secretion and nasal mucosa were taken from 23 patients with AR and 16 health individuals. The expression of RORC2 and IL-17 were detected by immunohistochemistry and quantitative real-time fluorescence reverse polymerase chain reaction. The allergic symptoms in patients were graded. RESULTS: The rate of positive cells of RORC2 and IL-17 in AR group were 0.17 ± 0.05 and 0.72 ± 0.13, higher than the 0.05 ± 0.02 and 0.27 ± 0.11 of health controls, the difference was statistically significant (t were 9.51 and 11.92 respectively, all P < 0.05). The expression level of RORC2 mRNA in nasal mucosa and peripheral blood of AR group were 0.063 ± 0.011 and 0.452 ± 0.031, higher than the 0.029 ± 0.009 and 0.239 ± 0.027 of health controls, the difference was statistically significant (t were 6.51 and 3.35 respectively, all P < 0.05). The concentrations of IL-17 in the nasal mucosa, nasal secretions and serum levels of AR group were (70.28 ± 10.69), (45.32 ± 8.55) and (6.76 ± 1.18) pg/ml, compared with (18.43 ± 8.34), (6.83 ± 1.31) and (0.74 ± 0.05) pg/ml of controls, the difference was statistically significant (t were 7.92, 17.66 and 15.43 respectively, all P < 0.05). The allergy symptom scores of AR group were 9.43 ± 1.27. There were correlations between the allergic symptom and the expression of RORC2 mRNA and IL-17 in nasal mucosa and peripheral blood (r value were 0.820, 0.746, 0.629, 0.841 respectively, all P < 0.05). CONCLUSION: RORC2 and IL-17 involved in the inflammatory response of AR and can be used as an indicator to judge the severity.

[Efficacy of sublingual immunotherapy with dermatophagoides farinae drops in patients with allergic rhinitis].

OBJECTIVE: To evaluate the efficacy of sublingual immunotherapy (SLIT) with dermatophagoides farinae drops in patients with allergic rhinitis (AR). METHODS: Two hundred and six patients with AR aged from 4 to 60 years were included in this study. Among them, 123 patients completed more than one year treatment, the number of patients completed one year, one year and a half, two years were 61, 41, 21 cases. Rhinitis symptom scores and medication scores of the three groups before and after treatment were compared. And rhinitis symptom scores and medication scores of patients with one year drug withdrawal after one and two years treatment were aslo compared. RESULTS: After SLIT one year, one year and a half, two years treatment, the symptoms in these patients were significantly improved compared with before. The symptom scores (x±s) were reduced from 6.00±2.27, 7.39±1.99 and 6.29±2.14 to 2.95±1.82, 3.28±2.58, 2.48±1.99. The differences were statistically significant (t value was 8.19, 10.29, and 5.97, all P<0.01). The proportion of patients without drug treatment of the three group were 68.9%, 73.2% and 80.9%, there was statistical significance before and after treatment in every group (value was 50.391, 43.619, 27.776, all P<0.01). Symptom improvement of sneezing, runny nose, nasal congestion, nasal itching were similar after one year, one year and a half, two years SLIT treatment, the differences were not statistically significant (F values were 1.200, 1.276 and 2.333, all P>0.05). The proportion of patients without drug treatment in the group which stopped medication one year after two year SLIT (76.2%) was higher than group stopped medication one year after one year SLIT (61.3%). There was no statistical significance (χ2=1.263, P>0.05). CONCLUSION: The dust mite drops can relieve symptoms after one year treatment, but the proposed two years treatment is important for the consolidation of improved symptoms, especially for the effect of reducing the use of symptomatic medication.