MiR-106a-5p modulates apoptosis and metabonomics changes by TGF-ß/Smad signaling pathway in cleft palate.

BACKGROUND: The molecular mechanisms of abnormal palatogenesis were investigated in this study. A key regulator, miR-106a-5p, and its target pathway were analyzed. OBJECTIVES: This research is trying to clarify the underlying mechanism of the modulation of miRNA transcription during the formation of cleft palate by 7T and 9.4T NMR metabolomic platforms. METHOD: Differentially expressed miRNAs and mRNAs were analyzed by microarray analysis and verified by qRT-PCR. The protein expression in TGFß signaling pathways were analyzed by Western Blotting. The relationship between miR-106a-5p and TGFß were analyzed by luciferase reporter assay. Cell apoptosis were analyzed by flow cytometer. And finally, the metabonomics were analyzed by NMR and multivariate data analysis models (MVDA). RESULTS: The expression of miR-106a-5p increased in cleft palatal tissue and negatively correlated with the protein level of Tgfbr2. The luciferase assay further proved that the tgfbr2 was a direct target of miR-106a-5p. In another aspect, miR-106a-5p increased apoptosis level in palatal mesenchymal cells, possibly because its inhibition of TGFß signaling pathway. Moreover, low cholesterol and choline levels with high citric acid and lipid levels were observed by 7T and 9.4T NMR metabonomic analysis, which inferred the disorder of cell membrane synthesis in cleft palate formation. Furthermore, transformation from choline to phosphatidylcholine regulated by miR-106a-5p was also disrupted, resulting in phosphatidic choline synthesis disorder and reduced cell membrane synthesis. CONCLUSIONS: The regulatory mechanism of cleft palate was studied at transcriptional and metabolomics levels, which may provide important information in understanding the primary cause of this abnormality.

Association between SATB2 gene polymorphism and cleft palate only risk in eastern Guangdong population and a meta-analysis.

To characterize the associations between the cleft palate (CPO) and single nucleotide polymorphisms (SNPs) of special AT-rich sequence-binding protein 2 (SATB2). We recruited 241 CPO and performed a case-control study with 242 controls. Concurrently, 103 of the patients and their normal parents were recruited to perform a case-parent trio study. Sixteen selected SNPs were genotyped. Furthermore, A meta-analysis was used to enhance the robustness of our conclusions. The case-control study provided no support for the hypothesis that any of the 16 selected SNPs played a significant role in CPO. In the meta-analysis, we also did not find that the SATB2 was associated with nonsyndromic cleft palate risk, in Asians or in Caucasians. The 16 selected SNPs do not contribute to the development of CPO.

Progress of epigenetic modification of SATB2 gene in the pathogenesis of non-syndromic cleft lip and palate.

Non-syndromic Cleft Lip and Palate (NSCLP) is one of the most common congenital craniofacial malformations. However, there is no enough knowledge about its mechanism, even through many relevant studies verify that cleft lip and palate is caused by interactions between environmental and genetic factors. SATB2 gene is one of the most common candidate genes of NSCLP, and the development of epigenetics provides a new direction on pathogenesis of cleft lip and palate. This review summarizes SATB2 gene in the pathogenesis of non-syndromic cleft lip and palate, expecting to provide strategies to prevent and treat cleft and palate in the future.

Association between Weight-Adjusted Waist Index and depressive symptoms: A nationally representative cross-sectional study from NHANES 2005 to 2018.

BACKGROUND: Depressive symptoms are strongly associated with the development of various diseases and are one of the leading causes of disability in the world. However, the relationship between weight-adjusted waist index (WWI) and depressive symptoms has not been studied. This study aimed to assess the relationship between depressive symptoms and WWI. METHODS: This study took NHANES data from 2005 to 2018 with 32,374 participants. Depressive symptoms were measured by a questionnaire (PHQ-9).WWI was determined by dividing the square root of waist circumference (cm) by weight (kg). Multivariate logistic regression models, smoothed curve fitting, and weighted generalized additive model (GAM) regression were used to examine the relationship between depressive symptoms and WWI, BMI, and waist circumference. Subgroup analyses and interaction tests were also performed. RESULTS: In fully adjusted models, the OR (95 % CI) for WWI and depressive symptoms with WWI, BMI, and waist circumference were 1.18 (1.05, 1.34), BMI 1.01 (1.00, 1.02, 1.01 (1.00, 1.01), respectively. Participants in the highest quartile (Q4) had a 49 % higher depressive symptoms compared to those in the lowest quartile (Q1) (OR = 1.49, 95 % CI:1.14-1.96). Subgroup analyses and interaction tests showed a stable relationship between depressive symptoms and WWI. LIMITATIONS: It is difficult to determine a causal relationship between the two; questionnaire collection may be somewhat biased; CONCLUSIONS: WWI was positively associated with depressive symptoms. This association was stronger than BMI and waist circumference. However, this relationship was stable. This study emphasizes the potential utility of WWI in preventing depressive symptoms and improving prognosis in the population.

Higher weight-adjusted waist index is associated with increased likelihood of kidney stones.

Objectives: The objective of this study was to evaluate the association between weight-adjusted waist index (WWI) and the prevalence of kidney stones among adults in the United States. Methods: The cross-sectional study utilized data from the National Health and Nutrition Examination Survey (NHANES) spanning the years 2007-2018. A total of 31,344 participants were categorized into two groups: those with kidney stones and those without. WWI was determined by dividing waist circumference (cm) by the square root of body weight (kg). To examine the relationship between kidney stones and WWI, multivariate logistic regression models, smoothed curve fitting, and weighted generalized additive model (GAM) regression were employed. Subgroup analysis and interaction tests were conducted to explore the stability of this association across different groups. Results: The final analysis comprised a total of 31,344 participants, including 2,928 individuals who had a history of kidney stones. In the fully adjusted model, an increase in WWI exhibited a positive correlation with the prevalence of kidney stones (OR=1.34, 95% CI: 1.18-1.51). When WWI was converted into quartiles (Q1-Q4), participants in the highest quartile (Q4) had a 69% greater risk of developing kidney stones compared to those in the lowest quartile (Q1) (OR=1.69, 95% CI: 1.28-2.25). This positive association was particularly notable among non-diabetic patients. Conclusion: Our study demonstrates a significant positive association between weight-adjusted waist index levels and an elevated prevalence of kidney stones among US adults. Furthermore, this research highlights the potential utility of weight-adjusted waist index in the prevention of kidney stones in the overall population. This relationship is limited and further research is needed to test this hypothesis.

Primary or metastatic branchial cleft carcinoma?: a case report.

The brachial cleft carcinoma is an extremely rare head and neck facial malignancy, and there is some disagreement about its differential diagnosis. In this paper, we report a 63-year-old male patient who had a mass on the left side of the neck and diagnosed as the brachial cleft carcinoma by intraoperative biopsy pathology. However, this patient was diagnosed with the carcinoma of the left soft palate more than 20 days after surgery and esophageal cancer 2 years later, and was treated accordingly. Therefore, it is hard to confirm whether the branchial cleft carcinoma is primary or metastatic. In fact, the diagnostic criteria for primary squamous cell carcinoma of branchial cleft cysts are very rigorous. Confirmation of the diagnosis is based on pathological examination of the branchial cleft cyst epithelium lined with squamous cells, meanwhile, a thorough examination should also be performed to exclude the presence of other primary cancers.

A LCMS-based untargeted lipidomics analysis of cleft palate in mouse.

BACKGROUND: Recent studies have shown that lipid metabolism was abnormal during the formation of cleft palate. However, the composition of these lipid species remains unclear. OBJECTIVE: Aims of this study were to identify the lipid species components and reveal the key lipid metabolic disorders in cleft palate formation. METHODS: The pregnant mice were divided into experimental group exposed to all-trans retinoic acid (RA-treated group) (n = 12) and control group (n = 12) at embryonic gestation day 10.5 (E0.5). The component of the palatal tissue metabolome was analyzed using a LCMS-based nontargeted lipidomics approach. Multivariate statistical analysis was then carried out to assess the differences between the RA-treated group and the control group. RESULTS: Twenty-nine lipid species were found to discriminate between RA-treated and control embryos. Among them, 28 lipid species increased and 1 lipid species decreased in the RA-treated group. Among these lipids, 13 were triglycerides, 9 were PEs, 3 were PCs, 2 were PSs, 2 were DGs. Further analysis of the number of carbons and unsaturated bond of triglycerides showed that TGs with high unsaturated bonds constituted a higher fraction in the RA-treated group. A higher amount of triglycerides containing 52, 54, 56, 58, 60 carbons, and 1 to 8 unsaturated bonds. Of note, under RA treatment, TG 50:1, 52:2, 56:6and 60:8 became the most prominent. CONCLUSION: Lipid metabolism is significantly different in the formation of cleft palate induced by RA, and the unsaturated triglycerides increased in the RA-treated group may play an important role in the formation of cleft palate.

Mouse embryonic palatal mesenchymal cells maintain stemness through the PTEN-Akt-mTOR autophagic pathway.

BACKGROUND: Both genetic and environmental factors are implicated in the pathogenesis of cleft palate. However, the molecular and cellular mechanisms that regulate the development of palatal shelves, which are composed of mesenchymal cells, have not yet been fully elucidated. This study aimed to determine the stemness and multilineage differentiation potential of mouse embryonic palatal mesenchyme (MEPM) cells in palatal shelves and to explore the underlying regulatory mechanism associated with cleft palate formation. METHODS: Palatal shelves excised from mice models were cultured in vitro to ascertain whether MEPM are stem cells through immunofluorescence and flow cytometry. The osteogenic, adipogenic, and chondrogenic differentiation potential of MEPM cells were also determined to characterize MEPM stemness. In addition, the role of the PTEN-Akt-mTOR autophagic pathway was investigated using quantitative RT-PCR, Western blotting, and transmission electron microscopy. RESULTS: MEPM cells in culture exhibited cell surface marker expression profiles similar to that of mouse bone marrow stem cells and exhibited positive staining for vimentin (mesodermal marker), nestin (ectodermal marker), PDGFRα, Efnb1, Osr2, and Meox2 (MEPM cells markers). In addition, exposure to PDGFA stimulated chemotaxis of MEPM cells. MEPM cells exhibited stronger potential for osteogenic differentiation as compared to that for adipogenic and chondrogenic differentiation. Undifferentiated MEPM cells displayed a high concentration of autophagosomes, which disappeared after differentiation (at passage four), indicating the involvement of PTEN-Akt-mTOR signaling. CONCLUSIONS: Our findings suggest that MEPM cells are ectomesenchymal stem cells with a strong osteogenic differentiation potential and that maintenance of their stemness via PTEN/AKT/mTOR autophagic signaling prevents cleft palate development.

Multi­organ assessment via a 9.4­Tesla MRS evaluation of metabolites during the embryonic development of cleft palate induced by dexamethasone.

The aim of the present study was to determine the association between maternal metabolism and development of the fetal palate, and to suggest a potential non­invasive prenatal diagnostic method for fetal cleft palate (CP). Dexamethasone (DXM) was used to create a CP mouse model. A 9.4­Tesla (T) magnetic resonance spectroscopy (MRS) imager was used to measure an array of metabolites in the maternal serum, placental tissue, amniotic fluid and fetal palates. Multivariate statistical analysis was performed using SIMCA­P 14.1 software. Following DXM treatment, variations were detected in multiple metabolites in the female mice and their fetuses based on 9.4T MRS. It was indicated that in the experimental group during CP formation, leucine, valine, creatine, acetate and citrate levels in the palatal tissue were lower, whereas lactate, alanine, proline/inositol and glutamate­containing metabolite levels were higher, compared with the levels in the control group. In placental tissue and amniotic fluid, succinate and choline levels were lower in the experimental group. The relative concentrations of cholesterol and lipids in palatal tissues from mice treated with DXM were higher compared with the concentrations in tissues from mice in the control group, with the exception of (CH2)n lipids. In the placental tissue, the alteration in cholesterol level exhibited the opposite trend. Lipid levels for the different lipid forms varied and most of them were unsaturated lipids.

Association Between an Interferon Regulatory Factor 6 Gene Polymorphism and Nonsyndromic Cleft Palate Risk.

Background: Involvement of interferon regulatory factor 6 (IRF6) gene polymorphisms in nonsyndromic cleft palate (NSCP) risk remains controversial. This investigation was performed to evaluate the relationship between IRF6 gene polymorphisms and NSCP risk. Materials and Methods: Two hundred forty-one patients with NSCP (including 103 complete trio families) were recruited, and 242 unaffected individuals were included as controls. Polymorphisms for the IRF6 rs2235371, rs801619, rs642961, rs44844880, and rs8049367 loci were characterized in both groups. Furthermore, eligible studies were identified from the databases through June 1, 2017, and were included in a meta-analysis to enhance the robustness of our conclusions. Results: The IRF6 rs2235371 A allele and AA genotype in the case group were found at higher frequencies than in the control group (A allele: p < 0.0016; AA genotype: p < 0.0049). The IRF6 rs801619 AA genotype and G allele were associated with NSCP risk (G allele: p < 0.0061; AA genotype: p < 0.0195). At the IRF6 rs642961, rs44844880, and rs8049367 loci genotype and allele frequencies were not statistically different between the NSCP group and normal controls. In the meta-analysis, the IRF6 A/G gene polymorphism (rs2235371) and IRF6 A/G gene polymorphism (rs642961) were associated with NSCP risk in the general population, whereas the IRF6 A/C gene polymorphism (rs2013162) was not. Conclusion: The IRF6 A/G gene polymorphisms at rs2235371 and rs642961, but not the IRF6 A/C gene polymorphism rs2013162, were associated with NSCP risk.

[Individualized surgical management of secondary rhinoplasty for cleft lip].

OBJECTIVE: ?To explore the effectiveness of transplantation of engraved autologous costal cartilage for individualized surgical management in secondary rhinoplasty for cleft lip. METHODS: ?Between September 2009 and January 2014, 350 patients with secondary nasal deformity of cleft lip were treated, including 160 males and 190 females with a mean age of 18.2 years (range, 16-56 years). Nasal deformity was caused by unilateral cleft lip in 200 cases and by bilateral cleft lip in 150 cases. The interval of lip repair and nasal deformity correction was 2-50 years (mean, 12 years). About a 2-6 cm cartilage was harvested from the 6th or 7th costal cartilage, and was engraved to the shape of "ge" in Chinese. The upper part was strengthened on both sides of the alar cartilage; the lower part was fastened to columella and nasal septum cartilages. The rest of cartilages was diced into 0.1 mm×0.1 mm×0.1 mm cubes. The columella incision was designed to "Z"-plasty, and was stretched to the nasion along alar edge. The engraved autologous costal cartilage was transplanted and fixed to the collapse of nostril. The cartilage cube was transplanted and filled into the collapse of nasal root to achieve the aesthetic effect of nasal augmentation. The effectiveness was evaluated according to the grade of secondary rhinoplasty for cleft lip by ZHANG Li et al. at 1, 6, and 12 months after operation. RESULTS: ?All incisions were primary healing. All patients were followed up 1-12 months (mean, 6 months). After rhinoplasty, the collapse of nostrils was lifted, and the shape and height of collapse of nostrils were symmetrical to normal side. The deflection of columella nasi was corrected. A beautiful shape of nose was achieved. The excellent and good rates were 98.6% at 1 month, 97.4% at 6 months, and 97.1% at 12 months after operation, showing no significant difference (χ2=0.545, P=0.761). CONCLUSIONS: ?The technique of transplantation of engraved autologous costal cartilage for individualized surgical management in secondary rhinoplasty for cleft lip can achieve excellent surgery effect.

Localization of Na(+)-K(+)-ATPase α/ß, Na(+)-K(+)-2Cl-cotransporter 1 and aquaporin-5 in human eccrine sweat glands.

In order to evaluate the function of the repaired or regenerated eccrine sweat glands, we must first localize the proteins involved in sweat secretion and absorption in normal human eccrine sweat glands. In our studies, the cellular localization of Na(+)-K(+)-ATPase α/ß, Na(+)-K(+)-2Cl-cotransporter 1 (NKCC1) and aquaporin-5 (AQP5) in eccrine sweat glands were detected by immunoperoxidase labeling. The results showed that Na(+)-K(+)-ATPase α was immunolocalized in the cell membrane of the basal layer and suprabasal layer cells of the epidermis, the basolateral membrane of the secretory coils, and the cell membrane of the outer cells and the basolateral membrane of the luminal cells of the ducts. The localization of Na(+)-K(+)-ATPase ß in the secretory coils was the same as Na(+)-K(+)-ATPase α, but Na(+)-K(+)-ATPase ß labeling was absent in the straight ducts and epidermis. NKCC1 labeling was seen only in the basolateral membrane of the secretory coils. AQP5 was strongly localized in the apical membrane and weakly localized in the cytoplasm of secretory epithelial cells. The different distribution of these proteins in eccrine sweat glands was related to their functions in sweat secretion and absorption.