OSMR gene effect on the pathogenesis of chronic autoimmune Urticaria via the JAK/STAT3 pathway.

BACKGROUND: Chronic autoimmune urticaria (CAU) is a common skin disease and remains unclear understanding of pathogenesis in the vast majority of cases. In order to explore a new therapy for CAU, the current study was performed to investigate the possible functioning of the Oncostatin M receptor (OSMR) gene in the autoimmunity of CAU via regulation of the JAK/STAT3 signaling pathway. METHODS: CAU skin tissues from 24 CAU patients and normal skin tissues from normal subjects were collected. Hematoxylin-eosin (HE) staining was conducted to count eosinophils, and immunohistochemistry was carried out to detect the positive rate of OSMR expression in two kinds of skin tissues. A total of 72 Kunming (KM) mice were selected, and 60 mice were used for establishing CAU models and later transfected with different plasmids. The expression of inflammatory factors was evaluated by enzyme-linked immunosorbent assays (ELISA). Expressions of janus kinase (JAK), signal transducer and activator of transcription 3 (STAT3), interferon-stimulated gene 15 (ISG15), CT10-regulated kinase (CRK), and interferon regulatory factor 9 (IRF9) were identified using Western blot assay and reverse transcription quantitative polymerase chain reaction (RT-qPCR). Epithelial cell proliferation was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, and cell cycle distribution and cell apoptosis were assessed using flow cytometry. RESULTS: The findings confirm that OSMR protein expression and histamine release rate are highly elevated in human CAU skin tissues, and the expression of the JAK/STAT3 signaling pathway-related genes (OSMR, JAK2, STAT3, ISG15, CRK and IRF9) was up-regulated. OSMR gene silencing in CAU mice significantly decreases the content of inflammatory factors (IL-1, IL-6, IFN-γ, and IgE), the number of eosinophils, and reduces the expression of the JAK/STAT3 signaling pathway related genes, and further enhances cell proliferation, promotes cell cycle entry and inhibits apoptosis of epithelial cells. CONCLUSION: All aforementioned results indicate that OSMR gene silencing inhibits the activation of the JAK/STAT3 signaling pathway, thereby suppressing the development of CAU.

A prospective study of topical carteolol therapy in Chinese infants with superficial infantile hemangioma.

BACKGROUND/OBJECTIVE: To report our observations from a trial of the short-term effectiveness and safety of topical carteolol hydrochloride drops to treat infantile hemangiomas (IHs). METHODS: From October 2012 to September 2015, the study recruited 349 children with superficial IHs. Participants were randomized to two groups: treatment (n = 224 who received 2% carteolol hydrochloride drops administered to the lesion surface twice daily) and observation (n = 125 who did not receive treatment). Therapy duration was 6 months. RESULTS: The mean age at the beginning of treatment was 3.2 months. Treatment responses were categorized as class 1 (total regression), class 2 (partial regression or controlled growth), or class 3 (no response). Of infants receiving carteolol treatment, 10.7% (24 patients) were categorized as class 1, 72.3% (162 patients) as class 2, and 17.0% (38 patients) as class 3. Of infants in the observation group, 5.6% (7 patients) were categorized as class 1, 25.6% (32 patients) as class 2, and 68.8% (86 patients) as class 3. No adverse effects were noted during treatment. CONCLUSION: Carteolol is an effective, safe topical treatment for superficial IHs. Carteolol may be used to treat proliferative superficial IHs, particularly in infants younger than 6 months.

[Be based on the morphological and histological changes to study optimal dose of TCDD induced cleft palate in mice embryo].

OBJECTIVE: To define the optimal 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) dose based on the morphological and histological changes of fetal mice cleft palate induced by different TCDD doses. METHODS: The pregnant mice were randomly divided into five groups and 6 in each grouop, and were gavaged on gestation day 10 (GD10). The control group were given 0.1 ml corn oil, and the experimental groups I, II , III, IV were given 32, 28, 24, 20 microg/kg TCDD respectively. To weight pregnant mice and embryos, record the number of live, cleft palate, dead and resorption fetal mice on GD 17.5. Another 15 pregnant mice were randomly divided into five groups (same as above) and 3 in each group. The coronal sections of the fetal mice heads were prepared at GD 13.5, 14.5 and 15.5 respectively, stained with haematoxylin-eosin staining (HE) and observed by microscopy. RESULTS: No significant differences in embryonic weight and live fetuses weight in each group. Compared with the control group,experimental groups I - III had small palate shelves (PS) and delayed palae shelves lift; the palate development and elevation in experimental group IV was similar to the control group. The incidence of cleft palate in the experimental groups I - IV were 97.37%, 93.02%, 65.12%, 56.82%, and no cleft palate in the control group. CONCLUSION: The optimal dose of TCDD to induce cleft palate in C57BL/6J mice is 28 microg/kg.

A retrospective study of propranolol therapy in 109 infants with infantile hemangioma.

The study reports the observations after propranolol therapy in 109 Chinese patients with infantile hemangioma. Response to treatment was favorable; 19 (17.4%) showed total regression, 89 (81.7%) partial regression, and 1 (0.9%) had no response. Twenty-three patients (21.1%) had some reactions, possibly due to the medication, but no life-threatening adverse effects were observed.

[The antagonistic effect of folic acid and resveratrol on cleft palate in mice induced by TCDD].

OBJECTIVE: To evaluate whether or not administration of folic acid and resveratrol have preventive effects on cleft palate formation as well as the comparison of the two drugs' s effects. METHODS: Pregnant mice were randomly divided into 9 groups, with 8 mice in each group. The TCDD group mice were dosed with TCDD 28 microg/kg body weight on gestation day 10 (GD 10) animals in folic acid group were respectively dosed with folic acid 15, 10, 5 mg/kg and TCDD 28 microg/kg; resveratrol treated mice were divided into 3 groups: resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13 in resveratrol (GD8-13 ) group; resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13, followed hy an oral administered with TCDD on GD10 in resveratrol (GD8-13) + TCDD group; resveratrol 50mg/kg and TCDD 28 microg/kg were used by gavage administration at GD10 in resveratrol (GD10) + TCDD group. Control mice were treated with the same volume of water for 6 consecutive days from GD8 to GD13 and were given a single dose of corn oil on GD10. The pregnant mice weight and embryos, the number of live, cleft palate, dead and resorption fetal mice were recorded on GD 17.5. The coronal sections of the fetal mice heads were prepared at GD 17.5 and observed by microscopy. RESULTS: Total frequency of clefts was 92.86% in TCDD group, 84.00% (15 mg), 73.08% (10 mg), 84.00% (5 mg) in folic acid + TCDD groups, 0% in resveratrol (GD10) group, 74.51% (GD10), 57.78% (GD8-13) in resveratrol + TCDD groups. The frequency of cleft was 0% in the control group. Compared with the control and the TCDD groups, there were significant differences in the number of live, dead and resorption fetal mice in TCCD + resveratrol (GD8-13) group (P < 0.05). No significant differences in embryonic weight, live fetuses weight, the number of live, dead and resorption fetal mice were found in the other groups (P > 0.05). CONCLUSION: Test dose of folic acid and resveratrol both had certain antagonistic effect on cleft palate in mice induced by TCDD, with folic acid 10 mg/kg, resveratrol 50 mg/kg GD8-13 doses having stronger antagonistic action. Effects of both the two drugs have no significant difference, but resveratrol (50 mg/kg, GD8-13) significantly affects the fetal mice's growth and development under TCDD exposure in utero.

[Mechanism of cleft palate in mice induced by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin].

OBJECTIVE: To explore the mechanism of cleft palate in mice induced by 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD). METHODS: On gestation day 10 (GD 10), 12 pregnant mice were randomly divided into two groups as the treated group and the control group with 6 mice in each group. The mice in the treated group received intragastric administration with 64 microg TCDD/kg, while the mice in the control group received equivalent corn oil. The embryos were examined under stereomicroscope to detect the incidence of cleft palate on GD 18.5. Another 18 pregnant mice were randomly divided into two groups (treated group and control group) on GD 10 with 9 pregnant mice in each group. Then each group was divided into 3 subgroups: GD 13.5, GD 14.5 and GD 15.5, with 3 pregnant mice in each subgroup. The palatal shelves were dissected from the embryos for RNA and DNA extraction on GD 13.5, GD 14.5 and GD 15.5. At last the expression of Smad 2-4 and Smad 7 mRNA was investigated by RT-PCR, and the TGF-beta3 promoter methylamine levels were investigated by methylation specific PCR (MSP). RESULTS: The cleft palate mice model was established successfully by exposing pregnant C57BL/6J mice to TCDD. Total frequency of clefts was 100% in TCDD group, and the frequency of clefts was 0 in the control group. The relative expression of Smad 2 mRNA was 0.263 +/- 0.088, 0.296 +/- 0.016 and 0.159 +/- 0.027 in TCDD group, 0.180 +/- 0.042, 0.282 +/- 0.029 and 0.165 +/- 0.018 in control group. The relative expression of Smad 3 mRNA was 0.453 +/- 0.153, 0.551 +/- 0.160 and 0.328 +/- 0.049 in TCDD group, 0.375 +/- 0.126, 0.510 +/- 0.145 and 0.259 +/- 0.035 in control group. The relative expression of Smad 4 mRNA was 0.675 +/- 0.174, 0.577 +/- 0.070 and 0.396 +/- 0.066 in TCDD group, 0.557 +/- 0.138, 0.587 +/- 0.080 and 0.441 +/- 0.054 in control group. The relative expression of Smad 7 mRNA was 0.283 +/- 0.050, 0.320 +/- 0.068 and 0.169 +/- 0.045 in TCDD group, 0.207 +/- 0.043, 0.288 +/- 0.051 and 0.155 +/- 0.040 in control group. There was no significant difference between the TCDD treated mice and the control (P > 0.05). The TGF-beta3 promoters were at the un-methylation state both in the TCDD treated and control group. CONCLUSION: It suggests that TCDD could induce a stable formation of cleft palate, but it is not through the TGF-beta/Smad signaling nor through the modification of TGF-beta3 promoter methylation.

Dynamic effect of di-2-(ethylhexyl) phthalate on testicular toxicity: epigenetic changes and their impact on gene expression.

This study investigated epigenetic (specifically, DNA methylation) changes and their impact on gene expression in testes induced by maternal exposure to Di-2-(ethylhexyl) phthalate (DEHP) in mice. Testicular dysgenesis syndrome was induced in fetuses and pups by maternal exposure to DEHP at 500 mg/kg/d, and testes were excised for analysis on gestation day (GD) 19 and postnatal days (PNDs) 3, 21, 56, and 90. High-performance liquid chromatography (HPLC) was performed to analyze DNA methylation status, and expression levels of the DNA methyltransferases were examined by quantitative real-time polymerase chain reaction (qPCR). Testis-specific gene, insulin-like hormone 3 (Insl3), and testosterone production were also detected. DEHP significantly increased DNA methylation levels on GD 19 and PND 3 (P < .05 and P < .05) but not on PNDs 21, 56, and 90. DEHP also significantly increased the expression of DNA methyltransferases. For DNA methyltransferase 1, the difference was not significant on PND 21, and DNA methyltransferase 3a and 3b returned to normal levels on PND 56. Fetal testes were a main target for DEHP as evidenced by a reduction in Insl3 expression and testosterone production. Effects of DEHP on Insl3 expression continued until PND 21. The DEHP-induced suppression of testosterone had not recovered on PND 56. Changes in DNA methylation may play an important role in abnormal testicular function caused by environmental factors such as maternal exposure to DEHP, which may be a mechanism of DEHP-mediated testicular toxicity.

Dynamic epigenetic changes involved in testicular toxicity induced by di-2-(ethylhexyl) phthalate in mice.

The aim of this study was to analyse epigenetic (specifically, DNA methylation) change in testes induced by maternal exposure to di-2-(ethylhexyl) phthalate (DEHP) in mice. Testicular dysgenesis syndrome was induced in foetuses by maternal exposure to DEHP. High-performance liquid chromatography was performed to analyse DNA methylation status, and expression levels of the DNA methyltransferases were examined by quantitative real-time polymerase chain reaction and western blotting. DEHP significantly had more than 10% relative increase in the global DNA methylation and also increased DNA methyltransferases' expression. Changes in DNA methylation may play an important role in abnormal testicular function caused by environmental factors such as maternal exposure to DEHP, which may be one possible mechanism of DEHP-mediated testicular toxicity.

Transforming growth factor-beta3 expression up-regulates on cleft palates induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) has been shown to induce cleft palate, in which the molecular etiology of the defect is poorly characterized. Recently, transforming growth factor-beta3 (TGF-beta3) has been indicated to play an essential role in the development of palatal shelves. In this developmental toxicity study, we investigated the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the expression of TGF-beta3 in fetal mice. Pregnant C57BL/6 mice were exposed to corn oil or TCDD (32 microg/kg/day 64 microg/kg/day, per os) at embryonic day 10 (ED10), a drastic inhibition of palatal shelves was induced. By using RT-PCR (reverse transcription-polymerase chain reaction) and Western blot, the expressions of TGF-beta3 was investigated. We found that the expression of TGF-beta3 was gradually up-regulated in TCDD-treated group. These results suggest that cleft palate can be induced by TCDD exposure, the modification of TGF-beta3 is related to its pathogenesis.

[Di-(2-ethylhexyl) phthalate increases the DNA methylation level of genomes in the mouse testis].

OBJECTIVE: To investigate the effect of the exposure to di- (2-ethylhexyl) phthalate (DEHP) during pregnancy on the DNA methylation level of genomes in the testis of the offspring in mice. METHODS: Pregnant KM mice were randomly divided into three groups, normal control, corn oil and DEHP-exposed. Corn oil and DEHP (500 mg/[kg x d]) were administrated respectively from gestation day 12.5 (GD 12.5) to postnatal day 3 (PND 3). The testes of the offspring were excised on PND 7, and their genomic DNA was treated with EcoR I /Msp I and EcoR I /Hpa II. The genome-wide DNA methylation patterns of the CCGG sites were detected by methylation-sensitive amplification polymorphism (MSAP). The samples were electrophoresed in the ABI 3730 DNA sequencer and the results analyzed by the Genescan3.1. RESULTS: The average incidence of DNA methylation was (34.03 +/- 3.05)% in the DEHP-exposed mice, obviously higher than (28.37 +/- 2.37)% in the normal control and (28.58 2.45)% in the corn oil group, with statistically significant differences (P < 0.05). CONCLUSION: Exposure to DEHP during pregnancy increases the DNA methylation level of the genome in the testis of the offspring and affects the apparent genetic modification of the genome, which may be one of the important toxicological causes of the lesion in the reproductive system.

[Oral mucosa flap in the top of fissure for nasal side mucosa repair of wide incomplete cleft palate].

OBJECTIVE: To explore a method to repair nasal side mucosa of wide incomplete cleft palate and reduce the tension of wound by using oral mucosa flap in the top of fissure. METHODS: 27 cases of wide incomplete cleft palatal were included in the study. On the basis of two-flap palatoplasty, the triangular oral mucosa flap in the top of fissure was turned and sewed with side mucosa to repair nasal side mucosa of wide palatal cleft. RESULTS: Without postoperative active bleeding, airway obstruction and wound infection, 27 cases had been repaired satisfactorily by this procedure. 1-3 months followed up demonstrated that all the wounds healed well without wound dehiscence or fistulas and the scars in the palate were not severe. CONCLUSION: Using oral mucosa flap in the top of fissure to repair nasal side mucosa of wide palatal cleft can get a reduced tension and correspondingly increase the width of mucoperiosteal flaps so as to decrease incidence rate of palatal fistulas and reduce formation of scars.

[Preliminary study of the ultrasonic measurement of thickness of skin in children].

OBJECTIVE: To collect the data of measuring skin thickness of children of both genders of different ages and parts of body with non-invasive high-frequency ultrasound method. METHODS: Two hundred and twenty-one children from 1 to 18 years of age,without systemic disease or injury in skin, were enrolled in the study and divided into 4 groups: i.e., infant group (112 years of age), pre-school age group (3-6 years of age), school age group (7-12 years for boys and 7-11 years for girls), adolescent age group (13-18 years for boys and 12-18 years for girls), and each group was subdivided into 2 groups according to the gender. The skin thicknesses of children in cheek, chest, abdomen, forearms, fundament and thigh was respectively measured by 13 MHz high-frequency ultrasound. RESULTS: The region with thinnest skin in children was the cheek, and the thickest was the back and buttock. (1) There were no significant differences in thickness of skin in the same region between genders and also among different age groups (P > 0.05). (2) There were also no obvious differences of thickness of the dermis and the whole skin in the same region between male and female, or among infants, pre-school age and school age groups (P > 0.05). In adolescent group, the average thickness of dermis in male was (1.16 +/- 0.04 ) - (1.98 +/- 0.47) mm, the average whole thickness of skin in male was (1.27 +/- 0.12) - (2.20 +/- 0.45) mm, while those of female were (1.00 +/- 0.18) - (1.60 +/- 0.30) mm and (1.10 +/- 0.17) - (1.83 +/- 0.29) mm (P < 0.05). CONCLUSION: It is reliable to measure the skin thickness by 13MHz ultrasound as a non-invasive method. The main factor which determined the thickness of the skin is dermal thickness, especially in males. The significant differences of skin thickness among cheek, back and buttock provide the basis for us to choose the appropriate thickness of skin grafts harvested from different body parts.