Serum apelin as a potential biomarker for infantile hemangiomas.

BACKGROUND: Infantile hemangiomas (IHs) are common benign vascular tumors in infants. Apelin, an endogenous cytokine, is implicated in the angiogenesis of neoplastic diseases. We aimed to explore the association between apelin and IHs, providing a foundation for clinical applications. METHODS: We identified differential expression of apelin in proliferative IHs compared to healthy controls (HCs) through bioinformatics analysis of publicly available databases and verified by Immunofluorescence. Enzyme-linked immunosorbent assay was used to quantify the serum levels of apelin and vascular endothelial growth factor (VEGF) in a cohort of 116 cases of proliferative IHs, 65 cases of capillary malformations (CMs), and 70 HCs. RESULTS: Apelin and APJ (APLNR, apelin receptor) were identified as the significantly upregulated differentially expressed genes (DEGs) in proliferative IHs. Immunofluorescence staining indicated high expression of apelin in proliferative IHs, while minimal expression in non-IH lesions. Apelin in IHs was reduced following 6 months of propranolol treatment. Serum apelin levels were significantly higher in the IH group compared to both the CM and HC groups. Moreover, apelin exhibited excellent discriminatory ability in distinguishing IHs from HCs, with an area under the curve (AUC) exceeding 0.90. A positive correlation was observed between the levels of apelin and the size of superficial IHs. The expression profiles of VEGF and apelin in IHs were found to be consistent. CONCLUSIONS: Apelin shows promise as a potential biomarker for IHs. The association between apelin and IH size, as well as its responsiveness to propranolol treatment, indicates its possible utility as a valuable indicator for the therapeutic evaluation of IHs.

Utilization of a Newly Designed Asymmetric Dorsal Gull Wing Flap for the Treatment of Postoperative Web Creep After Syndactyly Surgery.

BACKGROUND: Web creep is the most common long-term complication requiring revision after syndactyly surgery; however, few methods have been reported. The aim of this study was to introduce a newly designed asymmetric dorsal gull wing flap to reconstruct web for the postoperative web creep. METHODS: A retrospective analysis was performed for 20 patients from January 2016 to May 2019. Sex, age, original malformations, original surgical procedure, complications, time between the 2 operations, operation records, preoperative and postoperative photographs, and Withey score were reviewed. RESULTS: Eleven boys and 9 girls with average age of 60.65 ± 44.76 months underwent revision. Twenty-nine web spaces were affected (web creep, 12 cases; web creep and scar contracture, 17 cases). The original surgical procedure consisted of syndactyly separation in 15 cases, syndactyly separation with a full-thickness skin graft in 5 cases. There was 1 case of postoperative infection. All patients received an asymmetric dorsal gull wing flap and a zigzag incision, 15 patients received an additional full-thickness skin graft. The average time interval between the 2 operations was 34.60 ± 35.94 months. The follow-up time was 34.30 ± 20.73 months. No complications were noted, none of the patients redeveloped web creep. The median values for web creep, flexion-extension deformity, total Withey scores in the postoperative period were significantly lower than the preoperative values. The appearance and function of all digits were good. CONCLUSIONS: The asymmetric dorsal gull wing flap is a good choice for web reconstruction when web creep is caused by syndactyly surgery.

Polyotia: the Confusing Auricular Malformation.

OBJECTIVE: Polyotia is a very rare auricular malformation, and only few cases have been reported to date. Polyotia has been ambiguously defined, and due to the instability of its shape and condition, no uniform surgical technique has been established up to now. Thus, it is necessary to standardize the diagnosis and treatment of polyotia. The aim of the present study was to present a new set of objective diagnostic criteria for discussion, and introduce our surgical design for polyotia. METHODS: A retrospective analysis was performed on 34 cases of polyotia, which were diagnosed and treated in our Plastic Surgery Department during a 3-year period from January 2016 to March 2019. The preoperative photographs, manifestations and operation records of these 34 cases were reviewed. RESULTS: On the basis of the new set of objective diagnostic criteria, only 12 of 34 cases were diagnosed as polyotia, while the remaining 22 cases were diagnosed as accessory tragus. Polyotia was redefined as the presence of a broad-based accessory auricle in the tragus area along with accessory cavitas conchae similar to cavitas conchae. The new surgical design emphasized the use of cartilage and skin to fill up the concavity and reconstruct the tragus. CONCLUSIONS: The diagnosis of polyotia was presented on the basis of a new set of objective criteria, which include an accessory auricle and accessory cavitas conchae. The use of cartilage and skin to fill up the concavity and reconstruct the tragus were the emphases.

Dynamical Changes of Mandible and Upper Airway After Mandibular Distraction Osteogenesis in Pierre Robin Sequence.

OBJECTIVE: To present the dynamical evaluation of mandible and upper airway size among Chinese infant patients following mandibular distraction osteogenesis in a short-term follow-up and compare predistraction measurements with a normal age- and sex-matched control. METHODS: All the patients have undergone the computed tomography (CT) scan before mandibular distraction osteogenesis (T0), at the end of the distraction phase (T1), and 3 months after the end of the distraction phase before the distractor removal (T2). A CT analyzing computer software MIMICS was utilized to analyze the anatomic variables of upper airway size and mandible size. All analysis was based on a significance level of 0.05. RESULTS: Eight patients with Pierre Robin sequence differed mainly in the mandibular body length and the minimum anteroposterior dimension of the retroglossal airway from the control. After mandibular distraction osteogenesis, the mandibular body length and the ramus height both increased significantly, the ramus height also increased after 3 months of consolidation. Only small increase in the airway dimension of the retroglossal area at T2 was observed compared with T1. CONCLUSION: Mandibular distraction osteogenesis is an effective modality in treating Pierre Robin sequence. Compared with normal control, the main difference may be the length of mandibular body and the area of the retroglossal airway. There may not be an increase in the diameter of airway and the length of mandibular body after 3 months of growth and development in Pierre Robin sequence. Individual surgical plan should be made to gain a better prognosis.

Comparative Study of Folic Acid and α-Naphthoflavone on Reducing TCDD-Induced Cleft Palate in Fetal Mice.

OBJECTIVE: Tocompare the effect of folic acid (FA) and α-naphthoflavone on 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced cleft palate in fetal mice. DESIGN: Pregnant mice were randomly divided into seven groups. The mice treated with corn oil were used as a negative control. The mice in the other six groups were given a single dose of 28 µg/kg TCDD on GD 10 by gavage. For FA treatment, TCDD-treated mice were also dosed with 5, 10, and 15 mg/kg FA on GD 10, while for α-naphthoflavone treatment, the mice received a single dose of 50 µg/kg or 5 mg/kg α-naphthoflavone on GD 10. MAIN OUTCOME MEASURES: Fetal mice palates were imaged using light and scanning electron microscopy on GD 13.5, GD 14.5, and GD 15.5, and cleft palate were recorded on GD 17.5. The expression of guanosine diphosphate dissociation inhibitor (GDI) in fetal mice palate on GD 15.5 was examined by immunohistochemistry. RESULTS: TCDD successfully induced cleft palate. Ten mg/ml FA and 5 mg/ml α-naphthoflavone significantly reduced TCDD-induced cleft palate. FA and α-naphthoflavone partly reduced TCDD-induced cleft palate but did not affect the expression of Rho GDI. CONCLUSIONS: FA and α-naphthoflavone may reduce the generation of reactive oxygen species, inhibit MEE apoptosis through anti-oxidation, and increase filopodia and MEE movement. This may result in restoration of the ultrastructure of the palatal surface to a normal state, leading to the fusion and formation of complete palate in TCDD-treated fetal mice.

CENPF May Act as a Novel Marker and Highlight the Influence of Pericyte in Infantile Hemangioma.

Infantile hemangioma (IH), a benign microvascular tumor, is marked by early and extensive proliferation of immature hemangioma endothelial cells (Hem-ECs) that naturally regress through differentiation into fibroblasts or adipocytes. However, a challenge persists, as the unique biological behavior of IH remains elusive, despite its general sensitivity to propranolol treatment. Recent evidence suggests that abnormal volume proliferation in IH is primarily attributed to the accumulation of hemangioma pericytes (Hem-Pericytes), in addition to Hem-ECs. Centromere protein F (CENPF) is involved in regulating mitotic processes and has been associated with malignant tumor cell proliferation. It is a key player in maintaining genomic stability during cell division. Our findings revealed specific expression of CENPF in Hem-Pericytes, with a proliferation index (PI) approximately half that of Ki67 (3.28 vs 6.97%) during the proliferative phase of IH. This index decreased rapidly in the involuting phase (P < .05), suggesting that the contribution of pericytes to IH development was comparable to that of Hem-ECs. Tumor expansion and shrinkage may be due to the proliferation, reduction, and differentiation of Hem-Pericytes. In conclusion, we speculate CENPF as a novel marker for clinical pathological diagnosis and a potential therapeutic target, fostering advancements in drug development.

[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.

Early Wound Repair Versus Later Scar Repair in Children with Treadmill Hand Friction Burns.

The incidence of pediatric treadmill hand friction burns has been increasing every year. The injuries are deeper than thermal hand burns, the optimal treatment remains unclear. This was a retrospective study of children who received surgery for treadmill hand friction burns from January 1, 2015, to December 31, 2019, in a single burn center. A total of 22 children were surveyed. The patients were naturally divided into two groups: the wound repair group (13 patients), which was admitted early to the hospital after injury and received debridement and vacuum sealing drainage initially, and a full-thickness skin graft later; and the scar repair group (nine patients), in which a scar contracture developed as a result of wound healing and received scar release and skin grafting later. The Modified Michigan Hand Questionnaire score in the wound repair group was 116.31 ± 10.55, and the corresponding score in the scar repair group was 117.56 ± 8.85 (p > .05), no statistically significant difference. The Vancouver Scar Scale score in the wound repair group was 4.15 ± 1.21, and the corresponding score in the scar repair group was 7.22 ± 1.09 (p < .05). Parents were satisfied with the postoperative appearance and function of the hand. None in the two groups required secondary surgery. If the burns are deep second degree, third degree, or infected, early debridement, vacuum sealing drainage initially, and a full-thickness skin graft can obviously relieve pediatric pain, shorten the course of the disease, and restore the function of the hand as soon as possible.

TCDD induces cleft palate through exosomes derived from mesenchymal cells.

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) is a ubiquitous environmental toxicant and a notable teratogenic agent for cleft palate (CP), a common congenital structural malformation that can result from abnormalities during palatal shelf connection and/or fusion. The development of the palate requires precise coordination between mesenchymal and epithelial cells. Exosomes are vesicles secreted by cells and participate in organ development by transferring various bioactive molecules between cells and regulating cell proliferation, migration, apoptosis, and epithelial-mesenchymal transition (EMT); these vesicles represent a new method of intercellular communication. To explore how TCDD could influence palatal cell behaviors and communication, we treated mesenchymal cells with TCDD, collected the exosomes secreted by the cells, assessed the 2 types of palatal cells, and then observed the effects of TCDD-induced exosomes. We found that the effects of TCDD-induced exosomes were equal to those of TCDD. Thus, TCDD might change the genetic materials of palatal cells and exosomes to cause dysregulated gene expression from parental cells, affect cellular information communicators, and induce abnormal cellular behaviors that could lead to CP.

Protective effects of Radix Codonopsis on ischemia-reperfusion injury in rats after kidney transplantation.

PURPOSE: Kidney ischemia-reperfusion injury (IRI) after kidney transplant remains a major problem, separate from immune rejection that can lead to kidney transplant failure and graft function loss. Free radicals, disturbance of microcirculation and the inflammatory cascade appear to be the main contributors. Radix Codonopsis, a traditional Chinese drug used in vascular diseases, is an antioxidant and free radical scavenger. This study investigates the protective effect and underlying mechanisms of Radix Codonopsis extract saponins on kidney transplantation. METHODS: Renal transplantation was performed after rat kidneys had been stored for 1 h at 4°C. Blood urea nitrogen (BUN), serum creatinine (Scr), superoxide dismutase (SOD) and malondialdehyde (MDA) were assayed; bcl-2 and bax mRNA expression was detected using RT-PCR; bcl-2 and bax protein expression was detected by immunohistochemistry (IHC). Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) was used to detect apoptotic cells and determine the apoptosis index (AI). Analysis of variance (ANOVA) followed by Dunnett's test was used when more than two groups were compared. RESULTS: Saponin-treated animals showed increased SOD levels accompanied by decreased MDA, Scr and BUN levels (p < 0.05 vs. untreated controls); bcl-2 mRNA and protein levels were increased in transplanted kidney from treated animals, while bax mRNA and protein levels were decreased (p < 0.05 vs. untreated controls). AI was significantly decreased in transplanted kidneys from treated animals relative to untreated controls (p < 0.05 vs. untreated controls). CONCLUSION: This study clearly demonstrates the protective effects on IRI after kidney transplantation, which may be explained by decreased lipid peroxidation and inhibition of apoptosis.

Treadmill Hand Injuries Among Children: A Retrospective Case Series From Hospitalized Patients.

Background: With the progress of modernization, treadmill hand injury in pediatric population is taking on a global trend in recent years. The purpose of this study was to investigate the epidemiology and clinical features in a developing country, thereby providing some experience in the treatment and prevention of this particular type of injury. Methods: A 5-year retrospective review of patients with treadmill hand injury in Burn and Plastic Surgery ward at Children' Hospital of Chongqing Medical University was conducted. Demographics, injury details, therapy performed, length of hospital stay, complications, and outcome were analyzed. Results: Forty-six patients were surveyed, with a mean age of 3.5 ± 2.0 years old, including 24 males and 22 females. Injuries (77.8%) occurred between dinner to bedtime, and 95.7% happened indoors. Fingers were the most vulnerable part, of which the middle finger, ring finger, and index finger were the top three ones. The mean body surface area (BSA%) was 0.3 ± 0.2, but at least in deep dermal. Dressing changes, full-thickness skin grafts (FTSG), and Negative Pleasure Wound Therapy (NPWT) assisted FTSG were performed. The scar contracture, as the most severe complication, occurred in 26 patients, of which 22 originally received dressing changes at the time of injury. Conclusion: Treadmill hand injury in children should be highly regarded. Compared with conservative dressing changes, surgical intervention from a professional team may achieve more satisfactory prognosis and fewer complications. A prevention strategy based on "Time-Space-Person" was summarized according to its epidemiological characteristics, may help to decrease the incidence of this specific type of injury theoretically.

Anatomic Repair of Left Main Coronary Artery Atresia: Coronary Ostioplasty With Autologous Pulmonary Artery.

BACKGROUND: Left main coronary arterial (LMCA) atresia is a rare coronary arterial anomaly with extremely limited data on the optimal management. We aimed to report our single-surgeon experience of the ostioplasty in patients with LMCA atresia. METHODS: From July 2018 to December 2019, pediatric patients who presented with LMCA atresia and subsequently underwent surgical coronary ostioplasty were recruited into this retrospective study. Concomitant mitral repair was applied when the regurgitation was moderate or more severe. RESULTS: A total of 9 patients diagnosed with LMCA atresia were included. Mitral regurgitation was found in all of them, including 6 (66.7%) severe, 1 (11.1%) moderate, and 2 (22.2%) mild. In addition to ischemic lesions, which were found in 7 (77.8%) patients, structural mitral problems were also common (presented in 7 [77.8%] patients). All the patients underwent coronary ostioplasty with autologous pulmonary arterial patch augmenting the anterior wall of the neo-ostium. Mean aortic cross clamp time and cardiopulmonary bypass time was 88.1 ± 18.9 and 124.6 ± 23.6 minutes, respectively. During a median of 10.9 (range: 3.3 to 17.2) months' follow-up, there was only 1 death at 5 months after surgery. All survivors were recovered uneventfully with normal left-ventricular function; however, with 4 (50.0%) having significant recurrence of mitral regurgitation. CONCLUSIONS: With favourable surgical outcomes, coronary ostioplasty for LMCA atresia may be an option of revascularization. Structural mitral problems presented in majority patients, resulting in the requirement of concomitant mitral repair. However, the optimal technique of mitral repair remains unclear.

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.

A Case Series of Penile Skin Grafting in Children.

Pediatric penile skin grafting is rarely performed. We present a case series of four pediatric patients receiving skin grafting due to the loss of penile skin. The four boys were followed up for 1 to 5 years. One full-thickness skin graft and three split-thickness skin grafts (STSGs) survived well with low Vancouver scar scale scores. One boy gradually developed lymphedema of the distal foreskin and underwent a second preputioplasty. He presented with normal erectile function and did not experience any pain. We propose thick STSGs as the most appropriate choice for pediatric penile skin reconstruction. Lymphedema of the foreskin is an important long-term complication of penile skin grafting.

[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.

[Down-regulation of miR-381-3p inhibits osteogenic differentiation of mouse embryonic palatal mesenchymal cells in 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin-induced cleft palate of fetal mice].

OBJECTIVE: To investigate the correlation between down-regulation of miR-381-3p and inhibition of osteogenic differentiation of mouse embryonic palatal mesenchymal (MEPM) cells in 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD)-induced cleft palate of fetal mice. METHODS: Thirty-two pregnant mice were randomly divided into TCDD group and control group, 16 in each group. On embryonic day 10.5 (E10.5), the pregnant mice in TCDD group were orally administrated with TCDD at dosage of 28 µg/kg, while the pregnant mice in control group received equivalent corn oil. The pregnant mice in each group were sacrificed on E13.5 and E14.5, fetal palates were collected for analysis. The expression of miR-381-3p was detected by real-time fluorescent quantitative PCR and the protein expressions of runt- related transcription factor 2 (RUNX2) and osteopontin (OPN) were detected by Western blot. MEPM cells were extracted from fetal palates on E14.5 in control group and passaged. The 3rd passage cells were cultured with TCDD at dosage of 10 nmol/L for 0, 0.5, 1, 2, and 3 days. The expression of miR-381-3p was detected after 0, 0.5, 1, 2, and 3 days and the protein expressions of RUNX2 and OPN were detected after 0, 1, 2, and 3 days. Then, the 3rd passage cells were divided into 4 groups. The MEPM cells were transfected with miR-381-3p inhibitor (inhibitor group), NC inhibitor (NC inhibitor group) and miR-381-3p mimics (mimics group), NC mimics (NC mimics group) for 48 hours, respectively. And the expressions of miR-381-3p and the protein expressions of RUNX2 and OPN were detected. RESULTS: On E13.5 and E14.5, 96 fetal mice in control group and 92 in TCDD group were obtained. The bilateral palates contacted in control group on E14.5, and a gap between the bilateral palates existed in TCDD group. On E13.5 and E14.5, the relative expressions of miR-381-3p and RUNX2 and OPN proteins were significant lower in TCDD group than in control group ( P<0.05). The relative expression of miR-381-3p at 0.5 and 1 day after TCDD treatment of MEPM cells were significantly lower than that at 0 day ( P<0.05); then, the relative expressions at 2 and 3 days significantly increased, showing no significant difference when compared with that at 0 day ( P>0.05). The relative expressions of RUNX2 and OPN proteins at 1, 2, and 3 days were significantly lower than that at 0 day ( P<0.05). The relative expressions of miR-381-3p and RUNX2 and OPN proteins significantly lower in inhibitor group than in NC inhibitor group ( P<0.05) and higher in mimics group than in NC mimics group ( P<0.05). CONCLUSION: Down-regulation of miR-381-3p expression may be associated with inhibition of osteogenic differentiation of MEPM cells in TCDD-induced cleft palate of fetal mice.

Common differentially expressed proteins were found in mouse cleft palate models induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin and retinoic acid.

Cleft palate(CP) is a widely studied congenital malformation. However, its etiology and pathogenesis still remain unclear. Proteins are fundamental molecules that participate in every biological process within cells. In this study, we established CP mouse models induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and retinoic acid (RA), using proteomics technology isobaric tags for relative and absolute quantitation (iTRAQ) to investigate the key proteins in the formation of CP. Pregnant mice were given a gavage of TCDD 28µg/kg or retinoic acid 80mg/kg of body weight or equivalent corn oil at gestational day 10.5(GD10.5) and sacrificed at GD 17.5. Foetal mice were recorded and collected for further detection. Western blot was performed to verify the iTRAQ results. Eventually, we obtained 18 common differentially expressed proteins in TCDD group and RA group compared with normal control, 17 up-regulated and 1 down-regulated. 14-3-3sigma and Annexin A1 were up-regulated in experimental groups at GD17.5, which was consistent with Western blot. We speculated that the common differentially expressed proteins might be one of the molecular mechanisms in the formation of cleft palate.

The change in retinoic acid receptor signaling induced by prenatal marginal vitamin A deficiency and its effects on learning and memory.

This study investigated the prenatal marginal vitamin A deficiency (mVAD)-related impairment in learning and memory and the interactions between RARα, Src and NR1. Learning and memory were assessed in adult rats that were exposed to prenatal mVAD with Morris water maze. The average escape time was longer in mVAD rats, and they passed the hidden platform fewer times during the memory retention test than normal vitamin A intake (VAN) rats. The mRNA and protein levels of RARα, Src and NR1 in mVAD rats were significantly lower than those in VAN rats. RARα and Src, but not NR1, were in the same protein complex. RA treatment-induced increase in RARα, Src and NR1 expressions in mVAD neurons was much lower than that in VAN neurons. Overexpression of RARα gene in VAN neurons induced an increase in RARα, Src and NR1 expressions, while silencing of RARα gene induced a decrease in expressions of RARα and Src, but not that of of NR1. In mVAD neurons, however, overexpression of RARα did not induce an increase in NR1 expression, while silencing of RARα gene had no effect on Src and NR1 expressions. Furthermore, inhibition of Src was associated with a decrease in NR1 expression but not that of RARα. Prenatal mVAD was associated with impaired learning and memory in adult rats. It is possible that mVAD-related decrease in RARα led to a decrease in Src expression, which in turn down-regulated NR1 expression and Ca2+ influx and eventually caused learning and memory deficits.

Preliminary research on DNA methylation changes during murine palatogenesis induced by TCDD.

2,3,7,8-Tetrachlrodibenzo-p-dioxin (TCDD) has been shown to induce cleft palate through growth factor and receptor expression changes during palatogenesis. DNA methylation is an important epigenetic modification that can regulate gene expressions and may be involved in TCDD-induced cleft palate. In this study, we investigated the effects of TCDD on the global and CpG DNA methylation status and the expression levels of DNA methyltransferases (Dnmts) in palate tissue of fetal mice. Pregnant C57BL/6J mice were administered with corn oil or TCDD 28 µg/kg at gestation day 10.5(GD10.5), and sacrificed at GD13.5, 14.5, 15.5. Fetal palates were collected for molecular analysis. Global DNA methylation status was detected by Methylamp™ Global DNA Methylation Quantification Ultra Kit. The expression of DNA methyltransferases were examined by quantitative real-time PCR(q-PCR). Methylation Specific PCR (MSP) was performed to analyze CpG methylation status of Dnmts. We found that the global DNA methylation level and the expression of Dnmt3a were higher at GD13.5 in the TCDD group. The methylation level of CpG site 2 in the promoter region of Dnmt3a in the control group was higher than that of the TCDD group at GD13.5. The low CpG methylation level of Dnmt3a at GD13.5 which causes the up-expression of Dnmt3a may induce global hypermethylation in fetal palate tissue. The aberrant global methylation status at GD13.5 may be the cause of palate malformation in fetal mice induced by TCDD.

[Global DNA methylation changes during palatal formation in fetal mice induced by 2,3,7,8-tetrachlrodibenzo-p-dioxin].

Objective: To investigate global DNA methylation and DNA methyhransferases participation in the mechanism of cleft palate induced by maternal exposure to 2,3,7,8-tetrachlrodibenzo-p-dioxin (TCDD)in mice. Methods: 40 pregnant C57BL/6J mice were randomly divided into 2 groups: the control group(n =20) and TCDD-exposure group(n =20).On gestation day 10.5 (GD10.5),the mice in TCDD-group were orally administrated with TCDD 28 µg/kg, while the mice in the control group received equivalent corn oil. The pregnant mice were sacrificed on GD13.5,GD14.5,GD15.5,GD16.5,GD17.5,fetal palates were collected for analysis. Global DNA methylation levels were detected by Methylamp TM Global DNA Methylation Quantification Ultra Kit through an ELISA-like reaction. The expression levels of DNA methyltransferases were examined by quantitative real-time PC R(q-PCR).IBM SPSS 20.0 software was applied for statistical analysis. Kolmogorov-Smirnov test was used for normal distribution check, and the distribution was normal. Independent t-test was carried out among two groups. P ＜ 0.05 was considered statistically significant. Results: The global DNA methylation level in TCDD-exposure group was significantly higher than that in control group on GD13.5 (49.52％ ±4.03％ vs 33.42％ ± 6.78％,P ＜ 0.01),while lower on GD14.5 (24.10％ ±2.29％ vs 30.12％ ±3.92％,P ＜0.05) and on GD16.5 (32.77％ ±0.98％ vs 36.45％ ± 3.27％,P ＜ 0.05).The expression level of Dnmt1 mRNA in TCDD-exposure group was higher than that in control group on GD13.5(1.28±0.11 vs 1.01 ±0.10,P＜0.05) and on GD16.5(1.04 ±0.05 vs 0.81 ±0.01,P ＜0.01).The expression level of Dnmt3a mRNA in TCDD-exposure group was higher than that in control group on GD13.5 (1.15 ±0.17 vs 0.81 ±0.02,P ＜0.05)and on GD16.5 (1.11 ± 0.06 vs 0.96 ± 0.06,P ＜ 0.05).The expression level of Dnmt3b mRNA in TCDD-exposure group was higher than that in control group on GD14.5(0.97 ±0.06 vs 0.72 ±0.06,P ＜0.01). Conclusions: It is supposed that complicated mechanisms are exist to regulate global DNA methylation levels in palatal tissue of fetal mice. The significant increased DNA methylation level on GD13.5 resulting from up-expression of Dnmt1 and Dnmt3a may be one of the epigenetic mechanisms which cause palate malformation in fetal mice induced by maternal exposure to TCDD.