Corrigendum: Use of autologous cord blood mononuclear cells infusion for the prevention of bronchopulmonary dysplasia in extremely preterm neonates: a study protocol for a placebo-controlled randomized multicenter trial [NCT03053076].

[This corrects the article DOI: 10.3389/fped.2020.00136.].

[Three-dimensional finite element study on the effects of maxillary anterior two-step retraction and en-masse retraction with clear aligners].

Objective: To investigate the effects of two-step retraction and en-masse retraction on tooth movement pattern of anterior teeth and posterior anchorage with clear aligners using three-dimensional finite element analysis. Methods: A finite element model of maxillary first premolar extraction case undergoing clear aligner treatment was established based on maxillofacial cone-beam CT data of a 24-year-old adult male with individual normal occlusion, who visited Department of Oral Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine for impacted mandibular third molar in June, 2022. The initial tooth displacement of five anterior retraction protocols (two-step with canine retraction, two-step with incisor bodily retraction, two-step with incisor retraction-overtreatment, en-masse bodily retraction, and en-masse retraction-overtreatment) were evaluated. Results: Two step with canine retraction caused distal tipping of the canine and labial tipping of the incisors (0.18° for central incisor and 0.13° for lateral incisor). Two step with incisor retraction caused mesial tipping of the canine. In two step with bodily retraction protocol, uncontrolled lingual tipping was found in central incisor (0.29°) and lateral incisor (0.32°). In two-step with incisor retraction-overtreatment protocol, the movement pattern of the incisors didn't change, but the inclinations reduced to 0.21° and 0.18°. En-masse retraction caused distal tipping of the canine. In en-masse bodily retraction protocol, uncontrolled lingual tipping was also found in central incisor (0.19°) and lateral incisor (0.27°). In en-masse retraction-overtreatment protocol, the central incisor showed controlled lingual tipping (0.02°) and the lateral incisor showed palatal root movement (0.03° labial inclination). Posterior teeth exhibited mesial tipping in all five protocols. Conclusion: En-masse retraction with incisor overtreatment was beneficial to incisor torque control in clear aligner treatment.

[Effects of porcine urinary bladder matrix on motility and polarization of bone marrow-derived macrophages in mice].

Objective: To explore the effects of porcine urinary bladder matrix (UBM) on the motility and polarization of bone marrow-derived macrophages in mice, so as to provide evidence for the rational selection of stent in clinical wound repair. Methods: The method of experimental research was used. The microstructure of porcine UBM and absorbable dressing was observed under scanning electron microscope. Polyacrylamide gel electrophoresis was used to observe the protein distribution of the two stent extracts. The primary macrophages were induced from bone marrow-derived cells isolated from six 6-8-week-old male C57BL/6J mice (mouse age, sex, and strain, the same below) and identified. Three batches of macrophages were divided into porcine UBM extract group and absorbable dressing extract group. The cells in each group were cultured with Dulbecco's modified Eagle medium/F12 medium containing the corresponding extracts. The cell migration rate was detected and calculated on 1, 3, and 7 d after scratching by scratch test. The number of migrated cells at 12 and 24 h of culture was detected by Transwell experiment. The percentages of CD206 and CD86 positive cells at 24 h of culture was detected by flow cytometer. The numbers of sample in the above cell experiments were all 3. An incision was prepared on the left and right back of twelve mice, respectively. The left incision of each mouse was included in porcine UBM group and the right incision was included in absorbable dressing group, and the corresponding stents were implanted into the incisions respectively. On post operation day (POD) 7 and 14, the number of inflammatory cells infiltrated in the stent was detected by hematoxylin-eosin staining; the number of F4/80, transforming growth factor-ß1 (TGF-ß1), vascular endothelial growth factor (VEGF), and matrix metalloprotein-9 (MMP-9) positive cells and type Ⅰ collagen deposition in stents were observed by immunohistochemistry; the percentages of F4/80, CD86, and CD206 positive cells were observed by immunofluorescence staining. The numbers of sample in the above animal experiments were all 6. Data were statistically analyzed with analysis of variance for factorial design, analysis of variance for repeated measurement, and independent sample t test. Results: Porcine UBM has a dense basement membrane structure on one side and porous propria containing a fibrous structures on the other. Both sides of the absorbable dressing had three-dimensional porous structure. In the molecular weight range of (50-70)×103, multiple non-type Ⅰ collagen bands appeared in the lanes of porcine UBM extract, while no obvious bands appeared in the lanes of absorbable dressing extract. It had been identified that mouse bone marrow-derived cells had been successfully induced into macrophages. The cell migration rates in porcine UBM extract group were significantly higher than those in absorbable dressing extract group on 1, 3, and 7 d after scratching (with t values of 15.31, 19.76, and 20.58, respectively, P<0.05). The numbers of migrated cells in porcine UBM extract group were significantly more than those in absorbable dressing extract group at 12 and 24 h of culture (with t values of 12.20 and 33.26, respectively, P<0.05). At 24 h of culture, the percentage of CD86 positive cells in porcine UBM extract group ((1.27±0.19)%) was significantly lower than (7.34±0.14)% in absorbable dressing extract group (t=17.03, P<0.05);the percentage of CD206 positive cells in porcine UBM extract group was (73.4±0.7)%, significantly higher than (32.2±0.5)% in absorbable dressing extract group (t=119.10, P<0.05). On POD 7 and 14, the numbers of inflammatory cells infiltrated in the stents in porcine UBM group was significantly more than those in absorbable dressing group (with t values of 6.58 and 10.70, respectively, P<0.05). On POD 7 and 14, the numbers of F4/80, TGF-ß1, VEGF, and MMP-9 positive cells in the stents in porcine UBM group were significantly more than those in absorbable dressing group (with t values of 46.11, 40.69, 13.90, 14.15, 19.79, 32.93, 12.16, and 13.21, respectively, P<0.05); type Ⅰ collagen deposition in the stents in porcine UBM group was more pronounced than that in absorbable dressing group; the percentages of CD206 positive cells in the stents in porcine UBM group were significantly higher than those in absorbable dressing group (with t values of 5.05 and 4.13, respectively, P<0.05), while the percentages of CD86 positive cells were significantly lower than those in absorbable dressing group (with t values of 20.90 and 19.64, respectively, P<0.05), and more M2-type macrophages were seen in the stents in porcine UBM group and more M1-type macrophages were seen in the stents in absorbable dressing group. Conclusions: Porcine UBM can enhance macrophage motility, induce M2 polarization and paracrine function, create a microenvironment containing growth factors such as TGF-ß1 and MMP-9 tissue remodeling molecules, and promote tissue regeneration and extracellular matrix remodeling in mice.

[Clinical analysis of 49 cases of non-inflammasome related conditions].

Objective: To summarize the clinical characteristics and provide clues for early identification of non-inflammasome related conditions. Methods: The clinical manifestations, laboratory tests, genetic testing and follow-up of 49 children with non-inflammasome related conditions in Peking Union Medical College Hospital from January 2006 to February 2022 were retrospectively analyzed. Results: A total of 49 children, 29 of them were boys and 20 were girls. The age of onset was 0.8 (0.3, 1.6) years, the age at diagnosis was 5.7 (2.8, 8.8) years, and the time from onset to diagnosis was 3.6 (1.9, 6.3) years. Combined with genetic testing results, 49 children with non-inflammasome related conditions were found, including 34 cases (69%) of Blau syndrome, 4 cases (8%) of tumour necrosis factor receptor-associated periodic syndrome, 4 cases (8%) of haploinsufficiency of A20, 2 cases (4%) of Majeed syndrome, 2 cases (4%) of pyogenic sterile arthritis, pyoderma gangrenosum, acne syndrome and 3 cases (6%) of chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature syndrome. There were 22 cases (45%) with a positive family history. The clinical manifestations included 37 cases (76%) cases with rash, 38 cases (78%) with joint involvement, 33 cases (67%) with eye involvement, 17 cases (35%) with recurrent fever. In addition, 11 cases (22%) were complicated with digestive system involvement. Thirty cases (61%) presented as elevated inflammatory indexes (erythrocyte sedimentation rate and (or) C-reactive protein), positive autoantibodies were noticed in 3 cases (6%). The patients were treated with glucocorticoid in 23 cases (47%), immunosuppressive agents in 43 cases (88%) and biologic agents in 37 cases (76%). During the follow-up of 5.8 (2.9, 8.9) years, 3 cases (6%) died. Conclusions: The symptoms of non-inflammasome related conditions include recurrent fever, rash, joint and ocular involvement with increased inflammatory indexes and negative autoantibodies. Up to now, glucocorticoids, immunosuppressants and biologic agents are the most popular medications for the non-inflammasome related conditions.

[Clinical features and spinal lesions in patients with chronic prostatitis/chronic pelvic pain syndrome].

Objective: To analyze the clinical features and spinal lesions related to micturitionin of chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS) patients. Methods: Patients with CP/CPPS were enrolled to this study at the outpatient department of Tongji Hospital between January and June 2019. The data of clinical features was collected and analyzed, including lower urinary tract symptoms(LUTS), bowel syndrome and pain over different parts of body, as well as lower urinary tract dysfunction, spinal lesions and pelvic organ morphological changes demonstrated by MRI. The potential role of spinal lesions in the development of CP/CPPS syndrome was investigated. Results: A total of 126 CP/CPPS patients were included, with an age［M（Q1，Q3）］of 41(31,53) years and a course of disease of 2(1,20) years. Among them, 126 (100.0%) were complicated with LUTS, 72(57.1%) with bowel dysfunction and 88(69.8%) with pain. MRI showed the cervical central disc herniation(126 cases, 100.0%), the ischemic changing in the cervical area of visceral efferant pathway(82 cases, 65.1%), the lumbar central disc herniation(65 cases, 51.6%), and the sacral nerve cysts(97 cases, 77.0%) are commonly seen. In addition, the morphological changes in the visceral organs containing smooth muscle were demonstrated, including thickened bladder wall(91 cases, 72.2%), distended seminal vesicles(70 cases, 55.6%) and distended sigmoid colon/rectum(59 cases, 46.8%). Conclusions: CP/CPPS patients were characterized by the co-existence of LUTS, bowel dysfunction and somatic pain in one individual. The presence of multi-organ symptoms, combined with the high prevalence of spinal lesions associated with micturition reflex, suggesting the potential role of the spinal lesions in the development of CP/CPPS.

The extracellular vesicular pseudogene LGMNP1 induces M2-like macrophage polarization by upregulating LGMN and serves as a novel promising predictive biomarker for ovarian endometriosis recurrence.

STUDY QUESTION: How does ectopic endometrial stromal cell (Ecto-ESC)-derived extracellular vesicular Legumain pseudogene 1 (EV-LGMNP1), a newly identified pseudogene of Legumain (LGMN), contribute to M2-phenotype macrophage polarization, and does it predict recurrence in patients with ovarian endometriosis (EMs)? SUMMARY ANSWER: EV-LGMNP1, which is abundant in Ecto-ESCs and serum from ovarian EMs, can direct macrophages towards an M2 phenotype by upregulating LGMN expression and is a promising biomarker for predicting ovarian EMs recurrence. WHAT IS KNOWN ALREADY: Extracellular vesicles (EVs) can mediate cell-to-cell crosstalk to promote disease progression via cargo molecule transport. Recently, LGMNP1, a newly identified pseudogene of LGMN, has been reported to promote cancer progression by upregulating LGMN. LGMN is a well-studied protein that can induce M2-like polarization. STUDY DESIGN, SIZE, DURATION: An in vitro study was conducted with Ecto-ESCs isolated from ectopic endometrial samples, collected from two patients with ovarian EMs (diagnosed by laparoscopy and histological analysis). A clinical retrospective cohort study of 52 ovarian EMs patients and 21 controls with available preoperative serum samples was carried out (2013-2017). The follow-up period ended either at the time of recurrence or on 31 December 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS: Ecto-ESC-derived EVs (EV/Ecto-ESCs) were characterized by nanoparticle tracking analysis, transmission electron microscopy and western blotting. EV internalization by THP-1 cells, which are the most widely used primary human macrophages model, was detected by fluorescence labelling. After EV treatment, THP-1 cell polarization was detected by quantitative real-time PCR (qRT-PCR) and western blot analyses of CD86 (M1-related marker) and CD206 (M2-related marker). LGMNP1 mRNA expression level in EVs from both primary ectopic endometrioc stromal cells and serum was examined using qRT-PCR. Additionally, the expression of LGMN, the downstream target gene of LGMNP1, in THP-1 cells was evaluated using qRT-PCR and western blotting. Kaplan-Meier and multivariate Cox regression analyses were applied to evaluate the independent predictive factors of EMs recurrence-free survival. A novel nomogram model based on serum EV-LGMNP1 was then formulated to predict EMs recurrence. MAIN RESULTS AND THE ROLE OF CHANCE: In vitro assays demonstrated that EV/Ecto-ESCs drove macrophages towards an M2-like phenotype. Moreover, LGMNP1 contributed to EV/Ecto-ESC-induced M2 macrophage polarization by upregulating LGMN mRNA expression levels. Clinically, serum EV-LGMNP1 was more highly expressed in recurrent EMs patients than in controls and EMs patients without recurrence. Survival analysis and our novel nomogram reconfirmed that serum EV-LGMNP1 was a novel promising and meaningful non-invasive biomarker for predicting EMs recurrence. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: In vitro experiments were only performed on samples from two patients with ovarian endometriosis, and a larger sample size is needed. ESCs isolated from the eutopic endometrium of EMs and non-EMs patients should be studied in the future. Additionally, in vitro experiments should be performed using endometrial epithelium cells and further in vivo experiments, such as using mice endometriotic models to investigate whether EV/Ecto could induce M2 macrophage polarization, should be conducted. Moreover, multicentre, large-sample data are needed to validate our predictive nomogram model. WIDER IMPLICATIONS OF THE FINDINGS: Our study provides novel insights into the mechanism of M2 polarization involved in ovarian EMs progression mediated by an 'EV-shuttled pseudogene LGMNP1' mode. In addition, serum EV-LGMNP1 may serve as a novel non-invasive biomarker for predicting recurrence, providing a new therapeutic target for ovarian EMs. STUDY FUNDING/COMPETING INTEREST(S): This project was supported by funding from the National Natural Science Foundation of China (81971361), the Natural Science Foundation of Shanghai Science and Technology (19ZR1406900), the Shanghai 'Rising Stars of Medical Talent' Youth Development Program (AB83030002019004), the Clinical Research Plan of SHDC (SHDC2020CR4087), the Shanghai Municipal Health Commission (202040498), the Research and Innovation Project of the Shanghai Municipal Education Commission (2019-01-07-00-07-E00050) and the Clinical Research Plan of SHDC (SHDC2020CR1045B). There are no competing interests to declare.

[The value of serum glypican-3 level in aided diagnosis of patients with primary hepatocellular carcinoma].

Objective: To investigate the clinical value of glypican-3 (GPC3) detection in the diagnosis and therapy-monitoring of primary hepatocellular carcinoma (HCC). Methods: From March 2018 to May 2019, the patients with HCC were enrolled as the experimental group(n=166)from Fudan University Shanghai Cancer Centre, while the specimens from health control group(n=94) and benign control group (n=50) were analyzed. The serum of GPC3 and alpha fetoprotein (AFP)levels were respectively detected by ELISA and chemiluminescence. GPC3 detections combined with AFP etc. in accuracy of HCC diagnosis were explored by using Logistic regression analysis. Results: The serum GPC3 level in patients with HCC ［0.210 (0.048, 0.801)mg/L］ ［Median (quartile Q1, Q3)］ was significantly higher than those in healthy controls ［0.029(0.019, 0.052)mg/L］ and benign controls ［0.033(0.021, 0.043) mg/L］ (Z=-7.69, P<0.001).The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and AFP were significantly different among the three groups (Z=-7.02, -6.85, -8.36 respectively, P<0.001). Among the serological indicators, it was related to ALT and AST (Z=-3.77, -4.09 respectively, P<0.001).The Cut-off level of GPC3 was determined as 0.077 mg/L by ROC curve. The sensitivity of the combined detection of serum GPC3 with AFP for HCC was up to 87.82%, the specificity was 77.86%, the negative predictive value was 84.29%, and the positive predictive value was 82.53%.The HCC-GPC3 model was constructed by using Logistic regression analysis. The area under the ROC curve was 0.882, the total sensitivity was 91.10%, and the total specificity was 72.73%. Further analysis showed that the serum GPC3 of patients with HCC was significantly lower ［0.454(0.019, 0.286) mg/L］ than that before surgery［0.608(0.039, 0.554）mg/L］(Z=-7.32, P<0.001). Conclusion: The detection of serum GPC3 can be applied to aid diagnosis and therapy-monitoring of HCC.The combination of GPC3 and AFP can improve the diagnostic efficiency of HCC.

TFAP2A is a novel regulator that modulates ferroptosis in gallbladder carcinoma cells via the Nrf2 signalling axis.

OBJECTIVE: Ferroptosis is a recently identified form of controlled cell death generally associated with the accumulation of lipid-associated reactive oxygen species (ROS). However, the molecular mechanisms underlying ferroptosis have not been established. MATERIALS AND METHODS: Microarray expression data for three human gallbladder carcinoma (GBC) and matched non-tumour specimens were downloaded from the Gene Expression Omnibus (GEO) repository. Candidate genes were filtered using bioinformatic analysis. After cell transfection, candidate gene impacts on cell proliferation, migration, invasion and ferroptosis (ferrous iron (Fe2+) and malondialdehyde (MDA) levels) were assessed. RESULTS: We screened 626 differentially expressed genes (DEGs) including 465 that were downregulated and 161 that were upregulated in the three tissue pairs. These DEGs were used to construct a protein-protein interaction (PPI) network. Functional enrichment analysis revealed the top three modules in the network and four hub genes. Transcription factor AP-2 alpha (TFAP2A) was screened and showed overexpression in The Cancer Genome Atlas (TCGA) digestive system tumour data and a relationship with clinical survival. In vitro, GBC exhibited upregulated expression of TFAP2A, whose inhibition reduced GBC cell proliferation, migration, and invasion. Fe2+ and MDA levels were elevated. Moreover, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed TFAP2A enrichment in oxidative stress. Subsequent experiments demonstrated that TFAP2A silencing attenuated the expression of key genes associated with oxidative stress such as heme oxygenase 1 (HO-1), nuclear factor erythroid 2 like 2 (Nrf2), ferritin heavy chain 1 (FTH1) and NAD(P)H quinone dehydrogenase 1 (NQO1). CONCLUSIONS: Bioinformatic and experimental analyses reveal that TFAP2A plays a vital role in ferroptosis and hence is a potential therapeutic target for GBC treatment.

Use of Autologous Cord Blood Mononuclear Cells Infusion for the Prevention of Bronchopulmonary Dysplasia in Extremely Preterm Neonates: A Study Protocol for a Placebo-Controlled Randomized Multicenter Trial [NCT03053076].

Background: Despite the rapid advance of neonatal care, bronchopulmonary dysplasia (BPD) remains a significant burden for the preterm population, and there is a lack of effective intervention. Stem cell depletion because of preterm birth is regarded as one of the underlying pathological mechanisms for the arrest of alveolar and vascular development. Preclinical and small-sample clinical studies have proven the efficacy and safety of stem cells in treating and preventing lung injury. However, there are currently no randomized clinical trials (RCTs) investigating the use of autologous cord blood mononuclear cells (ACBMNC) for the prevention of BPD in premature infants. The purpose of this study is to investigate the effects of infusion of ACBMNC for the prevention of BPD in preterm neonates <28 weeks. Methods: In this prospective, randomized controlled double-blind multi-center clinical trial, 200 preterm neonates <28 weeks gestation will be randomly assigned to receive intravenous ACBMNC infusion (5 × 107 cells/kg) or placebo (normal saline) within 24 h after birth in a 1:1 ratio using a central randomization system. The primary outcome will be survival without BPD at 36 weeks of postmenstrual age or at discharge, whichever comes first. The secondary outcomes will include the mortality rate, other common preterm complication rates, respiratory support duration, length, and cost of hospitalization, and long-term outcomes after a 2-year follow-up. Conclusion: This will be the first randomized, controlled, blinded trial to evaluate the efficacy of ACBMNC infusion as a prevention therapy for BPD. The results of this trial will provide valuable clinical evidence for recommendations on the management of BPD in extremely preterm infants. Clinical Trial Registration: ClinicalTrials.gov, NCT03053076, registered 02/14/2017, retrospectively registered, https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S0006WN4&selectaction=Edit&uid=U0002PLA&ts=2&cx=9y23d4 (Additional File 2).

Sequential treatment by polidocanol and radiofrequency ablation of large benign partially cystic thyroid nodules with solid components: Efficacy and safety.

PURPOSE: The purpose of this study was to retrospectively evaluate the efficacy and safety of a sequential treatment including percutaneous polidocanol sclerotherapy and radiofrequency ablation (RFA) in terms of volume reduction and complication rate in large, benign, partially cystic thyroid nodules with solid components. MATERIALS AND METHOD: From April 2017 to April 2019, 46 patients with 47 large benign partially cystic thyroid nodules underwent sequential treatment. There were 14 men and 32 women with a mean age of 49.9±11.5 (SD) years (range: 18-75 years). The volume of initial nodules was 12.7±12.3 (SD) mL (range: 2.16-75.62mL). Volume reduction after percutaneous polidocanol sclerotherapy and further RFA was evaluated respectively. Patients had clinical and ultrasound evaluations at a follow-up time of 12.1±5.3 (SD) months (range: 1.5-23.9 months). Technical success and complications were accessed retrospectively. RESULTS: After unsatisfying results with polidocanol sclerotherapy alone the 46 patients with 47 large benign partially cystic thyroid nodules had further RFA. Mean volume reduction of 47 nodules was 90.5±11.3 (SD) % (range: 43.9-99.3%) one month after RFA, 94.9±6.2 (SD) % (range: 66.9-99.5%) three months after RFA, and 95.8±5.5 (SD) % (range: 71.0-99.8%) six months after RFA. No recurrence or nodule enlargement after RFA was observed at the last follow-up. The complication rate of RFA was 12.5% (8/46 patients), with minor complications only. CONCLUSIONS: The sequential treatment regimen, including percutaneous polidocanol sclerotherapy and RFA, is an appropriate and safe treatment strategy for large benign partially cystic thyroid nodules with solid components.

[rmIL-33-stimulated nuocytes promote allergic inflammation in mouse model of allergic rhinitis].

Objective:The study aimed to investigate the role of nuocytes in allergic rhinitis （AR） murine models. Method:After intranasal administration of recombinant （rm） interleukin （IL）-33 in BALB/c mice, nuocytes were sorted and purified from the mouse nasal-associated lymphoid tissue （NALT）. Then, we examined the response of nuocytes to rmIL-33 in vitro. After a murine model of AR was established using ovalbumin, we adoptively transferred the cultured NALT-derived nuocytes to mice models, and determined allergic responses in them. Result:rmIL-33 expanded nuocytes in NALT of mice compared with AR mice （t=3.66, P<0.01）, and increased production of IL-13 from these cells in vitro in comparison with unstimulated nuocytes （t=19.90, P<0.000 1）. After adoptive transfer of nuocytes, sneezing （t=9.89, P<0.000 1） ,numbers of eosinophils（t=8.17, P<0.000 1）, concentrations of IL-13 （t=40.47, P<0.000 1） and IL-33 （t=19.89, P<0.000 1） in nasal lavage fluid were all enhanced when compared with AR mice. Conclusion:Nuocytes promote allergic inflammation in a murine model of AR.

[The therapeutic effects of interleukin 10 gene-modified bone marrow-derived dendritic cells in a murine model of liver fibrosis].

Objective: To investigate the therapeutic effects and mechanisms of interleukin-10 (IL-10) gene-modified dendritic cells (DC-IL-10) in mice with liver fibrosis. Methods: DC-IL-10 was constructed in vitro, the phenotype and function of which were evaluated by flow cytometry. BALB/c mice were treated with intraperitoneal injection of carbon tetrachloride (CCl4) to establish liver fibrotic model. DC-IL-10 was administrated via tail vein. Animals were divided into 4 groups including normal dendritic cell(DC) control, liver fibrosis only, negative lentiviral transfection DC (DC-mock) and DC-IL-10. Liver function, cytokine secretion, T lymphocyte differentiation and liver histomorphology were tested. Real-time PCR and western blot were used to analyze the effect of DC-IL-10 on Wnt/ß-catenin signaling pathway and its role in liver fibrosis. Results: When compared with DC control and DC-mock, the expression of DC-IL-10 surface stimulating molecules (major histocompatibity complex-Ⅱ, CD(80), CD(86)) were significantly decreased (F=14.708, 22.503, 12.595, respectively, all P<0.05), and DC-IL-10 significantly inhibited T lymphocyte proliferation (F=50.295, P<0.05). When compared with liver fibrosis group, serum alanine aminotransferase and aspartate transaminase were decreased in DC-IL-10 treated group (all P<0.05), other parameters including inflammatory factors (tumor necrosis factor α, IL-6, IL-1ß) reduced (all P <0.05), the proportion of regulatory T cells (Treg) increased (F=6.742, P<0.05), pathological damage improved, the expression of Wnt3a, α-SMA and ß-catenin mRNA and protein significantly reduced in DC-IL-10 treatment group (all P<0.001) . Conclusions: DC-IL-10 induces elevation of Treg for immune tolerance, as well as inhibition of inflammatory response, block of Wnt/ß-catenin signaling pathway, which translates into improvement of liver fibrosis.

[Differentiation of naive T cells into Th2 cells induced by nuocyte cells in mice with allergic rhinitis].

Objective:The aim of this study is to examine whether nuocytes induced differentiation of primary T cells into Th2 cells in AR mice in vitro. Method:A murine model of AR on the background of BALB/c was established using ovalbumin, and nuocytes were sorted and purified from the mouse nasal associated lymphoid tissue (NALT) and cultured in vitro. Then, we assessed the expression of IL-4 in these cells. Mouse peripheral blood mononuclear cells were isolated and Th2 cells and T cells were isolated for in vitro culture, and the percentage of T cells in Th2 cells was detected. Then, NALT-derived nuocyte cells cultured in vitro were added to the above-mentioned mouse T cell culture medium for co-culture. Result:Numbers of sneezing, nasal rubbing and eosinophils in nasal lavage fluid were all enhanced in AR mice compared to normal ones. We used flow cytometry analysis to identify nuocytes from mice NALT as CD3CD4CD8CD19CD11bCD11cFcεR1 (lineage)-ICOS+, and also found that the cells expressed IL-4, and its protein and mRNA were all increased in AR mice versus normal mice. After nuocytes were co-cultured with T-cell cultures, we determined the percentage of Th2 cells in total T cells, and found that the percentage was increased significantly. Conclusion:nuocyte cells may induce the differentiation of primary T cells into Th2 cells in AR mice through IL-4 pathway.

[Nasal irrigation for the treatment of vasomotor rhinitis: a pilot study].

Objective: To observe the therapeutic effect of simple 3.0% saline nasal irrigation and combined treatment of 3.0% saline nasal irrigation and budesonide nasal spray for vasomotor rhinitis (VMR), and explore the long-term effect for VMR. Through examination of levels of substance P (SP) and mucin (MUC)5B in nasal lavage fluid, the mechanisms of nasal irrigation treatment for VMR was discussed. Methods: One hundred and one patients from Department of Otorhinolaryngology Head and Neck Surgery, Huashan Hospital of Fudan University with VMR were randomly divided into 4 groups. The number of patients was 24 in control group, 25 in budesonide nasal spray treatment group (budesonide group), 25 in nasal irrigation treatment group (nasal irrigation group) and 27 in budesonide nasal spray + nasal irrigation group (combined treatment group). Control patients were left untreated. Budesonide group was under budesonide nasal spray treatment, nasal irrigation group was treated using 3.0% saline with a temperature of 40℃ and combined treatment group was given both treatments. The duration of the intervention period was 3 months (90 days). Visual Analog Scale (VAS) was used to evaluate nasal symptoms, and the health-related quality of life was assessed using the 12-item Short Form Health Survey version 2.0 (SF-12v2). Enzyme-linked immunosorbent assay (ELISA) was used to assess the contents of SP and MUC5B in nasal lavage fluid before and after 3-month treatments in budesonide and nasal irrigation group in the study. MUC5B in nasal lavage fluid after the SP challenge and anticholinergic drug intervention in control group were also evaluated with ELISA. Results: Nighty out of 101 patients completed the study. In the budesonide and combined treatment group after relevant interventions, the total VAS score of nasal symptoms decreased (5.91±0.21 vs 3.82±0.15, 6.18±0.17 vs 3.92±0.15, t value was 8.193, 10.060, respectively, all P<0.05) and SF-12v2 score increased (146.00±1.23 vs 152.30±0.97, 146.00±1.08 vs 155.40±0.90, t value was 3.982, 6.697, respectively, all P<0.05), with both scores showed no significant differences in the nasal irrigation group (5.96±0.17 vs 5.72±0.15, 146.10±1.17 vs 147.00±0.94, t value was 1.038, 0.607, respectively, all P>0.05) after the first month. In the budesonide and combined treatment group after relevant interventions, the total VAS score of nasal symptoms decreased (5.91±0.21 vs 5.05±0.15, 6.18±0.17 vs 5.10±0.12, t value was 3.374, 5.351, respectively, all P<0.05) and SF-12v2 score increased (146.00±1.23 vs 150.90±0.76, 146.00±1.08 vs 153.60±0.94, t value was 3.373, 5.343, respectively, all P<0.05), with both scores showed no significant differences in the nasal irrigation group (5.96±0.17 vs 5.78±0.17, 146.10±1.17 vs 148.10±0.80, t value was 0.716, 1.438, respectively, all P>0.05) after the second month. By the end of the third month, in nasal irrigation and combined treatment group, the VAS score was diminished (5.96±0.17 vs 4.80±0.12, 6.18±0.17 vs 4.44±0.13, t value was 5.485, 8.264, respectively, all P<0.05) and SF-12v2 score was elevated (146.10±1.17 vs 150.80±0.96, 146.00±1.08 vs 152.90±0.85, t value was 3.163, 5.008, respectively, all P<0.05), but there were no significant differences in budesonide group (5.91±0.21 vs 5.68±0.18, 146.00±1.23 vs 148.40±0.85, t value was 0.819, 1.587, respectively, all P>0.05). Additionally, SP in nasal lavage fluid decreased and MUC5B showed no statistical changes in budesonide group after three months, however, SP showed no any changes and MUC5B reduced significantly in nasal lavage fluid in nasal irrigation group. Furthermore, the anticholinergic drug could not decrease the concentration of MUC5B after the SP challenge in nasal cavity in control group. Conclusions: The therapeutic effect of simple nasal irrigation with 3.0% saline or combined treatment of 3.0% saline nasal irrigation and nasal corticosteroids is superior to simple nasal corticosteroids. Nasal corticosteroids plays a role in the inhibition of sensory nerve endings in nasal mucosa, but neurotransmitter plays a limited role in the pathogenesis of VMR.

[Effect of BCYRN1 on proliferation and migration of airway smooth muscle cells in rat model of asthma].

Objective: To detect the effect of brain cytoplasmic RNA 1 (BCYRN1) on the proliferation and migration of airway smooth muscle cells (ASMCs) in rat model of asthma. Methods: Male SD rats were randomly divided into control group and asthma group (n=10 each). The ovalbumin (OVA) model was constructed in asthma group. Real time-qPCR was performed to detect the level of BCYRN1 in the ASMCs separated from the airway tissue of these rats. Then 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium (WST-1) assay, roche real-time cell analyzer assay and Transwell cell migration assay were performed to detect the viability/proliferation and migration of ASMCs which were transfected with Ad-BCYRN1.Platelet-derived growth factor (PDGF)-BB was used to treat ASMCs to induce proliferation and migration, and the level of BCYRN1 was examined.The viability/proliferation and migration of ASMCs treated with PDGF-BB and transfected with si-BCYRN1 were detected. Inspiratory resistance and expiratory resistance were measured in rats with BCYRN1 knockdown.Briefly, rats were randomly divided into four groups: control (group A), sensitization + Ad-GFP (group B), sensitization + AdSM22α-siBCYRN1 (group C), control + Ad-SM22α-siBCYRN1 (group D) (n=10 each). The corresponding adenovirus vectors were sent to lung of group B, group C and group D through nasal spray. The OVA model was constructed in group B and group C. The rats in group A and group D were treated with saline.After 24 h of the last treatment with OVA or saline, rats of each group were given tracheal intubation, connected with breathing machine. Rats were injected with methacholine to measure the inspiratory resistance and expiratory resistance. Results: The level of BCYRN1 in ASMCs separated from rats in asthma group and in ASMCs treated with PDGF-BB was 3.60±0.45 and 3.53±0.35, respectively, significantly higher than those of the corresponding control (both P<0.01). Ad-BCYRN1 significantly increased the expression of BCYRN1 in ASMCs. The cell viability and proliferation rates of ASMCs transfected with Ad-BCYRN1 increased 1.75-and 1.47-fold compared to those of the control group, respectively (P<0.01); mobility increased 2.42-fold compared to that of the control group (all P<0.01). BCYRN1 knockdown reversed the increasing proliferation and migration of ASMCs induced by PDGF-BB. The cell proliferation rate and cell migration number in the PDGF-BB treatment group were (4.87±0.21)% and 80.00±5.00, respectively, which were significant higher than those in the si-BCYRN1 transfected group ((3.63±0.21)% and 25.33±2.52, all P<0.01). BCYRN1 knockdown reduced the inspiratory resistance and expiratory resistance in sensitization + Ad-SM22α-siBCYRN1 group. When the concentration of acetylcholine reached 1 mg/kg, the inspiratory resistance in the group A, group B, group C, and group D were 8.27±0.21, 25.40±0.56, 12.07±0.67 and 8.40±0.46 cmH2O·s·ml-1, and expiratory resistance were 13.30±0.56, 38.37±1.33, 16.40±0.56 and 13.40±0.46 cmH2O·s·ml-1, respectively (all P<0.01). Conclusion: Overexpression of BCYRN1 promotes the proliferation and migration of ASMCs in rat model of asthma.

Investigation on the association between IL-10 C819T gene polymorphisms and susceptibility to gastric cancer.

We conducted a case-control study to investigate the association between the interleukin-10 (IL-10) C819T polymorphism and susceptibility to gastric cancer in a Chinese population. A total of 157 patients with gastric cancer and 249 controls were consecutively enrolled from the Guizhou Provincial People's Hospital between October 2012 and February 2015. The polymerase chain reaction-restriction fragment length polymorphism technique was used to genotype for IL-10 C819T. As determined by χ2-test, there was a significant difference in genotype distributions of IL-10 C819T between gastric cancer patients and controls (χ2 = 7.09; P = 0.03). Based on unconditional logistic regression analysis, the TT genotype of IL-10 C819T was significantly associated with increased risk of gastric cancer when compared with that of the CC genotype [odds ratio (OR) = 2.24; 95% confidence interval (CI) = 1.17-4.26; P = 0.008]. In a dominant model, we found that the CT + TT genotype of IL-10 C819T was associated with susceptibility to gastric cancer compared to that of the CC genotype (OR = 1.63; 95%CI = 1.02-2.64). In a recessive model, the TT genotype of IL-10 C819T was correlated with a higher risk of gastric cancer when compared with that of the CC + CT genotype (OR = 1.75; 95%CI = 1.01-3.02). In conclusion, our study suggests that the IL-10 C819T polymorphism is associated with an increased risk of gastric cancer in co-dominant, dominant, and recessive models.

Impaired balance of Th17/Treg in patients with nasal polyposis.

Nasal polyposis (NP) is a chronic inflammatory disease of the nasal cavity and sinuses that is regulated by T lymphocyte subsets. Imbalance of Th17/Treg has been considered critical in the development of inflammation and atopic reactions. To assess whether the balance of Th17/Treg is disrupted in patients with NP, we evaluated the distribution of Th17 and Treg cells among peripheral blood mononuclear cells (PBMCs) in atopic patients with NP, non-atopic patients with NP and controls. We then determined mRNA levels of RORc and Foxp3 and protein levels of IL-17, TGF-ß and IL-10 in polyp tissue among the three groups. Finally, we investigated the correlation between Th17-, Treg- and Th1-, Th2-related cytokines (INF-γ, IL-4, IL-5). The results demonstrated that both atopic and non-atopic patients with NP revealed significantly increased Th17 proportion and decreased Treg proportion in PBMCs, as well as significantly increased RORc and IL-17 levels and decreased Foxp3 and TGF-ß levels in polyp tissue. Furthermore, these differences were significant between atopic and non-atopic groups. The frequency of Treg in PBMCs was found to be negatively correlated with Th1 and Th2 cytokines in polyps. These results indicated that an impaired balance of Th17/Treg existed in patients with NP and was more severe in atopic patients, suggesting that the imbalance of Treg/Th17 may play an important role in the development of NP and that atopy may aggravate NP by promoting the imbalance of Th17/Treg.

RNA interference silences the human papillomavirus 6b/11 early gene E7 in vitro and in vivo.

BACKGROUND: RNA interference (RNAi) is a potential therapeutic tool in the treatment of various diseases, such as cancers and viral infections. Silencing of E7 genes is an effective method to suppress human papilloma virus (HPV)-related tumours. AIM: To determine the therapeutic potential of RNAi in controlling condyloma acuminatum (CA). METHODS: Small interfering (si)RNA duplexes or small-hairpin (sh)RNA-expressing plasmids targeting the E7 genes of HPV-6b or HPV-11were inoculated into cultured E7-expressing cells via cationic liposomes, or into E7 gene-expressing mouse tumour models intratumorally or intravenously. Experiments were performed in triplicate and E7 mRNA level was analysed by real-time PCR. RESULTS: The in vitro experiments found that both siRNAs and shRNA-expressing plasmids reduced the mRNA levels of HPV-6b or HPV-11 E7 to 20-40% at the optimum dosage of 25-50 nmol/L for siRNAs and 0.1-0.2 microg/mL for shRNA-expressing vectors. The optimum time for this to happen was 72 h. E7 mRNA expression in tumour models was reduced to 45-50% after three intratumural injections. Intratumoral injections of RNAi effectors induced greater inhibition than did intravenous injections. CONCLUSIONS: We conclude that HPV-6b/11 E7 gene expression can be specifically modulated by RNAi, which may provide a useful method in the management of CA.

Association of estrogen receptor alpha gene polymorphisms with bone mineral density in Chinese women: a meta-analysis.

INTRODUCTION AND HYPOTHESIS: A large number studies have examined the association between estrogen receptor alpha (ESR-alpha) gene polymorphisms and bone mineral density (BMD) in the Chinese population. We conducted a meta-analysis to assess their pooled effects. METHODS: We searched for all published articles indexed in MEDLINE, the Chinese Biomedical Database, and the Chinese Journal Full-text Database from January 1994 to April 2006. Any cross-sectional study that tested the association between ESR-alpha PvuII or XbaI genotypes and BMD at the femoral neck or spine in Chinese women was included in the review. Data were extracted independently by two reviewers using a standardized data extraction form. Sixteen eligible studies involving 4,297 Chinese women were identified. RESULTS: The overall frequencies of X and P alleles were 28% and 40%, respectively. The PvuII polymorphism was statistically significantly associated with BMD at the femoral neck (P = 0.038 for PP = Pp = pp) but not at the lumbar spine in all women. The BMD difference for the contrasts of PP versus Pp/pp genotypes was -0.0105 (95%CI, -0.0202 approximately -0.0008) g/cm(2) (P = 0.036). The XbaI polymorphism was not associated with BMD at the femoral neck or lumbar spine. CONCLUSION: The PvuII polymorphism had a very weak association with femoral neck BMD whereas XbaI polymorphism was unlikely to be a predictor of femoral neck or spine BMD in Chinese women.

Formation of MTBE-DNA adducts in mice measured with accelerator mass spectrometry.

Methyl tert-butyl ether (MTBE) is a gasoline oxygenate and antiknock additive substituting for lead alkyls currently in use worldwide. Previous studies have shown that MTBE at very high doses induces tumors in rodents. The aim of the present study was to examine directly the binding ability of MTBE onto DNA, demonstrating its potential genotoxicity. MTBE-DNA adducts and their decay kinetics in mice have been measured by using doubly 14C-labeled MTBE with an advanced, ultrasensitive technique: accelerator mass spectrometry (AMS). It was found that MTBE definitely formed adducts with DNA in mouse lung, liver, and kidney in a log/log linear dose-response relationship. The distribution sequence of DNA adducts in these tissues is: lung > liver > kidney. The level of MTBE-DNA adducts peaked at 12 h postadministration in the lung and peaked at 6 h postadministration in the liver. Then the adducts declined rapidly until 5 days postadministration and thereafter declined much more slowly. To our knowledge, this is the first report on DNA adduction with MTBE in vivo. The mechanism of the formation of MTBE-DNA adducts also is discussed.