Saikosaponin-A Exhibits Antipancreatic Cancer Activity by Targeting the EGFR/PI3K/Akt Pathway.

BACKGROUND: A diagnosis of pancreatic cancer is pretty grim. Saikosaponin-A (SSA) is a Chinese herbal extract with anticancer activity. However, the therapeutic effect of SSA on pancreatic cancer remains elusive. AIM: The study aims to evaluate the antitumor effects of SSA on pancreatic cancer cells in vitro and in vivo. METHODS: After treatment with SSA, cell viability was measured using the CCK-8 assay, DAPI staining was performed to analyze the effect on nuclear morphology, propidium iodide (PI) staining was used to detect the cell cycle, and Annexin V/PI double staining was conducted to analyze apoptosis. Then, the expression of apoptosis-related proteins and EGFR/PI3K/Akt pathway-related proteins was determined using western blotting. The binding of SSA to EGFR was analyzed by performing molecular docking. The mouse pancreatic cancer model was established by subcutaneously injecting pancreatic cancer cells, and after 30 days of SSA gavage, the tumor volume was calculated. Tumor tissue sections were subjected to Ki67 immunohistochemical staining and HE staining. RESULTS: SSA inhibited the proliferation of pancreatic cancer cells. As the concentration of SSA increased, the proportions of BxPC-3 and MIA PaCa-2 cells in the G0/G1 phase increased, the proportions of early and late apoptotic cells also increased, and the apoptosis rate gradually increased. Apoptosis inhibitor experiments indicated that SSA promoted the activation of caspase 3 to induce apoptosis in pancreatic cancer cells. In addition, SSA treatment significantly reduced the levels of phosphorylated EGFR, Akt, and PI3K in the two cell lines. Molecular docking results showed that SSA may have potential binding sites in EGFR. Results of the xenograft experiment confirmed the antitumor effects of SSA, as evidenced by the decreased tumor weight and downregulated expression of Ki67. CONCLUSION: The results revealed that SSA exerted inhibitory effects on pancreatic cancer cells. These effects may be related to the inactivation of the EGFR/PI3K/Akt signalling pathway.

Reoperation for Recurrent and Persistent Cushing's Disease without Visible MRI Findings.

PURPOSE: Transsphenoidal surgery is the first-line treatment for Cushing's disease (CD), even with negative preoperative magnetic resonance imaging (MRI) results. Some patients with persistent or recurring hypercortisolism have negative MRI findings after the initial surgery. We aimed to analyze the efficacy of repeat surgery in two groups of patients and determine if there is an association between positive MRI findings and early remission. PATIENTS AND METHODS: Clinical, imaging, and biochemical information of 42 patients who underwent repeat surgery by a single neurosurgeon between 2002 and 2021 was retrospectively analyzed. We compared the endocrinological, histopathological, and surgical outcomes before and after repeat surgery among 14 CD patients with negative MRI findings and 28 patients with positive MRI findings. RESULTS: Immediate remission was achieved in 29 patients (69.0%) who underwent repeat surgery. Among all patients, 28 (66.7%) had MRI findings consistent with solid lesions. There was no significant difference in remission rates between the recurrence and persistence groups (77.8% vs. 57.1%, odds ratio = 2.625, 95% confidence interval = 0.651 to 10.586). Patients in remission after repeat surgery were not associated with positive MRI findings (odds ratio = 3.667, 95% confidence interval = 0.920 to 14.622). CONCLUSIONS: In terms of recurrence, repeat surgery in patients with either positive or negative MRI findings showed reasonable remission rates. For persistent disease with positive MRI findings, repeat surgery is still an option; however, more solid evidence is needed to determine if negative MRI findings are predictors for failed reoperations for persistent hypercortisolism.

Autosomal recessive monilethrix: Novel variants of the DSG4 gene in three Chinese families.

BACKGROUND: Monilethrix is a rare hereditary hair loss disorder characterized by hair fragility and beaded hair shaft alterations. Monilethrix is classically inherited in an autosomal dominant (AD) fashion caused by variants in the hair keratin genes KRT81, KRT83, or KRT86. Interestingly, an autosomal recessive (AR) form of monilethrix with variants in DSG4 gene has also been reported in recent years. OBJECTIVE: To identify causative variants in Chinese patients with autosomal recessive (AR) form of monilethrix. METHODS: Three families with AR form of monilethrix were observed and sequence variant analysis of DSG4 was performed by polymerase chain reaction (PCR), quantitative real-time PCR, and DNA sequencing. RESULTS: All the patients had sparse, fragile hair involving the scalp, eyebrows, and eyelashes with keratotic follicular papules and pruritus since birth. Atypical-beaded hairs and broken hair shaft fragments were identified in all the patients under dermoscopy. Heterozygous variants c.837del and c. 2389C > T, a homozygous splice site variant c.2355 + 1G > A, and a homozygous 48,644 bp large deletion variant g.31381440_31430084del in the DSG4 gene were identified and verified in the families. CONCLUSION: This report provided further evidence for the phenotypic spectrum and clinical features of, and the expanded variant database of AR form of monilethrix.

Case report of neurofibromatosis type 1 combined with primary ciliary dyskinesia.

Neurofibromatosis (NF) is a genetic disease in which the lungs are rarely involved. However, in NF cases with lung involvement, chest computed tomography may show bilateral basal reticulations, apical bullae, and cysts without bronchiectasis. Herein, we report a patient diagnosed with NF on the basis of the results of genetic testing who presented with early-onset wet cough and bronchiectasis. Considering the differential diagnosis of bronchiectasis combined with his early-onset wet cough, sinusitis, and sperm quality decline, we considered the possibility of primary ciliary dyskinesia (PCD). Further electron microscopy analysis of cilia and identification of homozygous mutations in the RSPH4A gene confirmed the diagnosis of PCD. Therefore, for patients with NF, when an image change exists in the lungs that does not correspond to NF, the possibility of other diagnoses, including PCD, must be considered.

The microarray identification circular RNA hsa_circ_0105015 up-regulated involving inflammation pathway in essential hypertension.

BACKGROUND: Essential hypertension (EH) is an inflammatory disease, and endothelial dysfunction induced by chronic inflammation is one of the pathogeneses of EH. The expression of some inflammatory mediators may be regulated by the interaction of circular RNAs (circRNAs) and microRNAs (miRNAs). METHODS: An Agilent human circRNA microarray was used to identify the expression profile of circRNAs in EH. qRT-PCR was used to evaluate the relative expression of circRNAs in 48 pairs of human whole blood samples (sex and age ± 3 years matched) and endothelial cells. TNF-α was applied to induce endothelial cells inflammation. CircRNA-miRNA network was predicted by MiRanda software. RESULTS: There were 287 circRNAs differentially expressed in the microarray. The top 10 up-regulated circRNAs in the EH group were hsa_circ_0014243, hsa_circ_0133228, hsa-circRNA14116-3, hsa_circ_0079536, hsa-circRNA13649-1, hsa_circ_0117886, hsa_circ_0007075, hsa-circRNA15285-1, hsa-circRNA10088-9, and hsa-circRNA14119-10; the top 10 down-regulated circRNAs were hsa_circ_0100094, hsa_circ_0127342, hsa_circ_0093773, hsa_circ_0096334, hsa_circ_0131618, hsa_circ_0063886, hsa_circ_0097804, hsa_circ_0126640, hsa-circRNA8935-1, and hsa_circ_0039978 (fold change in descending order). Hsa_circ_0105015 has two predicted binding sites with hsa-miR-637. The relative expression of hsa_circ_0105015 in EH patients was significantly higher than healthy controls (P = .002), and similar results appeared in TNF-α-induced endothelial cells. The area under the curve after hsa_circ_0105015 combined with hsa-miR-637 was 0.703, P < .001. CONCLUSION: Hyperexpression of hsa_circ_0105015 is a significant risk factor of EH and its association with EH involves inflammatory pathways. Hyperexpression of hsa_circ_0105015 combined with hypoexpression of hsa-miR-637 indicates vascular inflammation or endothelial dysfunction and has potential as a biomarker for early diagnosis of EH.

Hsa_circ_0037897 may be a risk factor for essential hypertension via hsa-miR-145-5p.

Objectives: This study aims to investigate the association between hsa_circ_0037897 and essential hypertension (EH) and to evaluate the diagnostic value of biomarker hsa_circ_0037897 in EH. Methods: This study included 92 EH patients and 92 sex- and age- (±3 years) matched subjects as control. qRT-PCR was performed to measure the expression level of circRNA and miRNA. Logistic regression analysis model was used to assess independent association between hsa_circ_0037897 and EH. Results: The expression level of hsa_circ_0037897 in EH patients was significantly higher (p < .001) compared to the control group, while hsa-miR-145-5p had significantly lower expression(p = .002) than the control group. The area under the ROC curve (AUC) of hsa_circ_0037897 was 0.656. Furthermore, the AUC increased to 0.714 when hsa_circ_0037897 was combined with hsa-miR-145-5p, BMI and smoking. Conclusion: The present results suggested that the high expression of hsa_circ_0037897 may be a risk factor for EH, and hsa_circ_0037897 has certain diagnostic value for EH.

The up-regulated hsa-circRNA9102-5 may be a risk factor for essential hypertension.

BACKGROUND: The present study was aimed to investigate the expression levels of circular RNAs (circRNAs) in the peripheral blood of essential hypertension (EH) patients and healthy controls (HC). On this basis, we tried to explain the possible role of circRNAs in the progression of EH and their potential as diagnostic biomarkers of EH. METHODS: First, we analyzed the differentially expressed circRNAs in peripheral blood obtained from the finished microarray analysis and selected four circRNAs under strict standards. Then, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure the expression levels of the selected circRNAs in a total of 192 blood samples, consisting of 96 HC and 96 diagnosed EH patients. Bioinformatics prediction of the target microRNAs (miRNAs) was performed for differentially expressed circRNAs, and the circulating vascular-related miRNAs were selected for qRT-PCR analysis to determine their expression levels. RESULTS: Hsa-circRNA9102-5 (11.7 ± 1.06 vs 12.13 ± 1.11, P = .007) was up-regulated in the patients group which was diagnosed with EH, as compared to the HC group, and was involved in the regulation of EH by sponging hsa-miR-150-5p. The area under the ROC curve (AUC) of the model was 0.620, using hsa-circRNA9102-5 as an independent predictor. Furthermore, the AUC was increased to 0.728 when hsa-circRNA9102-5 was combined with hsa-miR-150-5p and multiple other factors, as a combined predictor. CONCLUSIONS: The present results suggested that hsa-circRNA9102-5 may have played a crucial role in the development of EH by sponging hsa-miR-150-5p, which showed great potential as a novel target.

A potential risk factor of essential hypertension in case-control study: MicroRNAs miR-10a-5p.

Background: Essential hypertension is a multifactorial disease with high morbidity. The researches on the influence of genes on the disease are still in its infancy, and the mechanism of gene regulation is not clear. MiRNAs are key molecules that regulate the expression control of protein-coding or protein-non-coding RNA. It may be an important biological molecule risk factor for essential hypertension.Methods: A case-control study with 98 EH and 98 non-EH was conducted in our experiment. The candidate miRNAs including miR-10a-5p and miR-497-5p were detected and verified by qRT-PCR.Results: The expression level of miRNA in EH cases was significantly lower than the healthy control (P = 0.005). In addition, the relative expression of miR-10a-5p was closely positive correlated with DBP (r = 0.162, P = 0.023) and SBP (r = 0.223, P = 0.002). After adjusting confound factors, the result of the logistic regression indicated that hypo-expression of miR-10a-5p is a risk factor for EH (OR(95%CI) = 1.676(1.302,2.157), adjusted P < 0.0001). And the ROC analysis shows that the combined line with BMI and miR-10a-5p was a values marker for EH (AUC: 0.728, P < 0.0001).Conclusions: Lower expression of miR-10a-5p, as the key role, is significantly related to the risk of EH and maybe as a potential biomolecule for EH.

Clinicopathological analysis of IgA nephropathy combined with other glomerular diseases.

It is not rare to find Immunoglobulin A (IgA) nephropathy (IgAN) combined with other glomerular diseases, which can be called compound IgAN (cIgAN). Till now, clinical-pathological investigation of cIgAN was lacking, especially the severity of "background IgAN lesions." This research aimed to investigate the incidence, clinical and pathological characteristics of cIgAN, and thus improve the understanding of the clinical significance of this combination.Patients with cIgAN diagnosed in Peking University People's Hospital from November 2012 to April 2018 were retrospectively analyzed. Patients with IgAN without compound glomerular diseases (sIgAN) were enrolled as a control group.Among 1407 patients diagnosed with IgAN, 80 (5.69%) were cIgAN patients. Compared with sIgAN, cIgAN patients had a significantly lower prevalence of microscopic hematuria and more urine protein. There were 10 pathological types of glomerular diseases combined with IgAN, led by diabetic nephropathy 37 (46.25%) and membranous nephropathy 14 (17.5%). Histologically, although the mesangial hypercellularity was comparable in 2 groups, cIgAN patients had a lower prevalence of endocapillary proliferation, segmental glomerulosclerosis, and cellular or fibrocellular crescents formation, as well as weaker immunofluorescence intensity for IgA and C3 (all P < .05). Eight out of 27 (29.63%) cIgAN patients with follow-up data (5-48 months) developed irreversible end-stage renal disease requiring dialysis.The order of incidence of concomitant diseases was similar to that of the pure diseases. The "background IgAN associated lesions" except mesangial hypercellularity were relatively mild in cIgAN group. Those might suggest that in some cases, IgAN seems to be a chance finding, and the combined diseases may play a more important role in the clinicopathological features of the patients than the nephritis caused by IgA deposition. While diagnosing IgAN, other combined glomerular diseases need to be carefully considered by nephrologists and pathologists.

Microarray Profiling of Circular RNA Identifies hsa_circ_0126991 as a Potential Risk Factor for Essential Hypertension.

Essential hypertension (EH), a major cause of cardiovascular diseases, is an important public health issue. However, the molecular mechanisms involved in EH remain unknown. Circular RNA (circRNA) is a novel promising biomarker for the disease. The purpose of the present study was to determine the expression of circRNAs in the blood of EH patients and to evaluate the performance of circRNA for early diagnosis of EH. A total of 178 subjects were recruited in the case-control study. Initial screening was done by using the Agilent human circRNA microarray followed by qRT-PCR validation. Finally, miRNAs were combined with circRNAs to create a new early prediction model for EH. The expression level of hsa_circ_0126991 in EH patients was significantly higher in comparison with healthy controls (p < 0.0001). Using the interaction of miR-10a-5p in combination with hsa_circ_0126991 led to a sensitivity of 0.708, a specificity of 0.764, and combined area under the curve of 0.774 (95% CI: 0.705-0.843) for early diagnosis of EH. In summary, the present study uncovered a novel perspective that hyperexpression of hsa_circ_0126991 is correlated with the risk of EH and may serve as a stable biomarker for early diagnosis of EH.

Up-regulation of circular RNA hsa_circ_0037909 promotes essential hypertension.

AIMS: Essential hypertension (EH) is a high prevalence disease facing a public health challenge. People were little known about the genetics of diagnosing the cause of EH. Circular RNAs that have a continuous cycle of covalent closure, without affected by RNA exonuclease, and are more stable and hard to degrade may involve into the molecule regulation mechanism of EH as an important biomedical. METHODS: qRT-PCR was used to analyze circRNAs in total volume of human blood and the induced human aortic endothelial cells (HAECs) and human umbilical vein endothelial cells (HUVECs). Our case-control study was involved with 48 pairs of case controls with sex and age (±3 years) match. We conducted t test, Pearson's χ2 test, and receiver operating characteristics (ROC) curve analysis for the corresponding analysis. RESULTS: The expression level of hsa_circ_0037909 in EH patients was significantly higher than that in the healthy controls (P = 0.007), and the expression level of hsa-miR-637 in EH patients was significantly lower in than that in the healthy controls (P = 0.039); the same result appears in the HAECs and HUVECs. Hsa-miR-637 (adjusted P = 0.018), hsa_circ_0037909 (adjusted P = 0.005), HDL (adjusted P = 0.024), and serum creatinine (adjusted P = 0.014) were brought into the model which performed logistic regression analysis. The combination of two RNAs was excellent (P < 0.001) through ROC curve analysis. Hsa_circ_0037909 was significantly positively correlated with serum creatinine (P < 0.001) and low-density lipoprotein (LDL) (P = 0.017). CONCLUSIONS: Our findings suggested that the combination of hsa_circ_0037911 and hsa-miR-637 may be a significant important biomarker for early diagnosis of EH. Hsa_circ_0037909 may affect serum creatinine or LDL leading to the formation of EH.

Circular RNA hsa_circ_0014243 may serve as a diagnostic biomarker for essential hypertension.

Circular RNAs (circRNAs) have a great potential as clinical biomarkers; however, specific circRNAs with a diagnostic value for essential hypertension (EH) largely remain to be identified. In the present study, the potential application of Homo sapiens (hsa)_circ_0014243, which was identified to be significantly upregulated in whole blood samples of EH patients in a previous microarray profiling study by our group, in the diagnosis of EH was evaluated. Reverse transcription-quantitative polymerase chain reaction analysis was performed to determine the expression levels of hsa_circ_0014243 and hsa-microRNA (miR)-10a-5p in a total of 178 blood samples collected from 89 healthy controls and 89 patients diagnosed with EH. Divergent primers were designed for circRNAs, while conventional primers were used for miRs. Independent-samples t-tests and bivariate correlation analyses were performed to analyze the association between clinical factors influencing EH and hsa_circ_0014243 expression levels. A receiver operating characteristics (ROC) curve was generated to estimate the diagnostic value of hsa_circ_0014243 for EH. Finally, the expression levels of circRNAs and miRNAs were combined to propose a possible prediction model for EH. The results indicated that hsa_circ_0014243 was upregulated in whole blood samples of EH patients compared with that in the controls (P<0.001). Furthermore, the relative expression levels of hsa_circ_0014243 (Δ quantification cycle) were identified to be significantly correlated with age (r=-0.259, P<0.001), high-density lipoprotein levels (r=0.196, P=0.009) and glucose levels (r=-0.204, P=0.006). The area under the ROC curve (AUC) of the model using hsa_circ_0014243 as a predictor was 0.732. Of note, the AUC increased to 0.781 when hsa_circ_0014243 levels were combined with hsa-miR-10a-5p levels as predictors. The present results suggest that hsa_circ_0014243 has a crucial role in the genesis and development of EH, and presents a certain diagnostic capability for EH.

Hypomethylation of the Interferon γ Gene as a Potential Risk Factor for Essential Hypertension: A Case-Control Study.

Essential hypertension (EH) is a multifactorial disease. Interferon-γ (IFN-γ) plays an important role in the onset of EH through cytokine-mediated systemic inflammatory responses. We aimed to determine whether the methylation status of the IFN-γ gene (IFNG) promoter is involved in the pathogenesis of EH. Six copies of CpG dinucleotides are distributed between 3,203 bp and 3,121 bp upstream from the transcription initiation site of IFNG, termed CpG1 to CpG6 in the 5'-to-3' direction. We recruited 96 patients with EH and 96 sex- and age-matched healthy subjects as controls. Using bisulfate pyrosequencing datasets, we analyzed the methylation status of the six CpG sites and thus found that CpG5 was consistently methylated in all of the 96 EH patients and 96 control subjects. Among the remaining five CpG sites, there was no significant difference in the methylation levels of CpG4 and CpG6 between the two groups. By contrast, CpG1 (P = 0.003) and CpG3 (P = 5.87 × 10-7) were highly methylated among the EH subjects compared with the controls, whereas CpG2 (P = 1.24 × 10-12) was significantly less methylated in among EH subjects. The methylation levels of CpG2 were still lower after adjustment with logistic regression (adjusted P = 0.032). The CpG2 methylation level was an effective marker of EH (area under curve = 0.384; P = 1.40 × 10-15). The present study shows that hypomethylation of the IFNG promoter is significantly related to the risk of EH, providing new insights into the pathogenesis of EH.

A potential risk factor of essential hypertension in case-control study: Circular RNA hsa_circ_0037911.

BACKGROUND: Essential hypertension (EH) is a high prevalence with multifactorial diseases. Human studies on the impact of genes on this disease are just in the initial stage, the mechanism of gene regulation is still remains unclear. Circular RNAs (circRNAs) as a continuous cycle of covalent closure, RNA molecules added to the 3'-5' end covalently bound by the formation of incidental event. CircRNAs may be an important biomolecule in revealing the molecule regulate mechanisms of EH. METHODS: The circRNAs were selected and validated with qRT-PCR followed. Our experiment was conducted with case-control studies among 200 EH participants. The t-test was used to evaluate the different expression of circRNAs and miRNAs, the significance of which was set as p < 0.05. RESULTS: The hsa_circ_0037911 expression level in EH cases were significantly higher than healthy controls (p = 0.005). There was still important significance when adjusted by logistic regression (adjusted p = 0.026). We also found that hsa_circ_0037911 was an effective marker of EH (area under curve = 0.627; p = 0.002). The levels of hsa_circ_0037911 were significantly differences in gender, BMI, smoking and drinking among EH cases. There was a positive correlation between Serum creatinine (Scr) and hsa_circ_0037911. CONCLUSIONS: Our findings suggested that higher expression hsa_circ_0037911 may be key circRNAs for EH development by changing the concentration of Scr and could be a stable biomarker for early diagnosis of EH.

RIP140/PGC-1α axis involved in vitamin A-induced neural differentiation by increasing mitochondrial function.

Vitamin A deficiency and mitochondrial dysfunction are both associated with neural differentiation-related disorders, such as Alzheimer's disease (AD) and Down syndrome (DS). The mechanism of vitamin A-induced neural differentiation and the notion that vitamin A can regulate the morphology and function of mitochondria in its induction of neural differentiation through the RIP140/PGC-1α axis are unclear. The aim of this study was to investigate the roles and underlying mechanisms of RIP140/PGC-1α axis in vitamin A-induced neural differentiation. Human neuroblastoma cells (SH-SY5Y) were used as a model of neural stem cells, which were incubated with DMSO, 9-cis-retinoic acid (9-cis-RA), 13-cis-retinoic acid (13-cis-RA) and all-trans-retinoic acid (at-RA). Neural differentiation of SH-SY5Y was evaluated by Sandquist calculation, combined with immunofluorescence and real-time polymerase chain reaction (PCR) of neural markers. Mitochondrial function was estimated by ultrastructure assay using transmission electron microscopy (TEM) combined with the expression of PGC-1α and NEMGs using real-time PCR. The participation of the RA signaling pathway was demonstrated by adding RA receptor antagonists. Vitamin A derivatives are able to regulate mitochondrial morphology and function, and furthermore to induce neural differentiation through the RA signaling pathway. The RIP140/PGC-1α axis is involved in the regulation of mitochondrial function in vitamin A derivative-induced neural differentiation.

Metabolic Profiling of Plasma in Gestational Diabetes Mellitus Using Liquid Chromatography and Q-TOF Mass Spectrometry.

BACKGROUND: To delineate the metabolomic profiling and identify early diagnostic biomarkers in maternal plasma from the pregnant women who subsequently developed gestational diabetes mellitus (GDM) using liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-Q-TOF MS). METHODS: Plasma samples were collected from GDM pregnant women (n = 30) and healthy controls (n = 30) at the 20th gestational week in Huzhou Central Hospital and Huzhou Maternal and Child Health Hospital. The principle component analysis (PCA), partial least-squares discriminant analysis (PLS-DA), and orthogonal PLS (OPLS) were sequentially applied to discover the differential metabolites for GDM diagnosis. Further, we analyzed the identified biomarkers in the MetPA database in order to reveal the key relevant metabolism in GDM. RESULTS: Twenty-four out of 975 aligned metabolites were distinguished among GDM plasma and healthy controls. In particular, the level of linolenic acid and arachidonic acid was significantly elevated in GDM. CONCLUSIONS: The linolenic acid and arachidonic acid could be selected as new potent biomarkers for GDM diagnosis and prognosis in early pregnancy; however, they still need to be confirmed from large samples in future.

Comparison of the safety of electrotome, Harmonic scalpel, and LigaSure for management of thyroid surgery.

BACKGROUND: Energy-based surgical devices, including electrotome, the Harmonic scalpel, and LigaSure, have been widely applied in thyroid surgery, although a comparison of their safety and efficacy has not been reported yet. In this study, we investigated the feasibility of using hemostatic energy-based surgical devices during thyroid surgery in a canine model. METHODS: Twenty-four beagle dogs were randomly divided into the following groups: electrotome (30 kW), electrotome (15 kW), the Harmonic scalpel (output level 3), and LigaSure (middle gear). The hemostatic devices were applied on the thyroid surface for 3 seconds and then near the recurrent laryngeal nerve (RLN; distance of 5 mm, 3 mm, or 1 mm) for 3 seconds. Evoked electromyography (EMG) amplitudes were recorded by intraoperative neuromonitoring (IONM). Acute microstructural morphological damage to thyroid tissues and the RLN were evaluated immediately after the procedure by light and electron microscopy. RESULTS: Electrotome caused a significant decrease in evoked EMG amplitudes when applied at a vertical distance of 1 mm from the RLN, both at 30 kW (1046 ± 404.3 µV vs 153 ± 245.5 µV; p < .001) and 15 kW (1197 ± 589.2 µV vs 986.3 ± 797.3 µV; p = .037), compared with those evoked under normal conditions. Furthermore, distinct acute microstructural morphological changes of the RLNs were observed by light and electron microscopy. However, no significant functional or histological changes were induced by the electrotome at a vertical distance of 5 mm or 3 mm from the RLN. The Harmonic scalpel and LigaSure induced neither marked changes in evoked EMG amplitudes when applied at vertical distances of 5 mm, 3 mm, or 1 mm (all p > .05) nor microstructural morphological changes in the RLNs. The electrotome (15 kW) caused more serious thermal damage to thyroid tissues than that caused by either the Harmonic scalpel or LigaSure (thermal damaged depth: 0.951 ± 0.061 vs 0.756 ± 0.074, p < .001; 0.951 ± 0.061 vs 0.724 ± 0.116, p < .001). Nevertheless, there were no differences between the Harmonic scalpel and LigaSure groups (p = .435). CONCLUSION: LigaSure and the Harmonic scalpel might be safer than electrotome when used in thyroid operations. LigaSure generates less heat than the Harmonic scalpel and electrotome. © 2017 Wiley Periodicals, Inc. Head Neck 39: 1078-1085, 2017.

A bama minipig model of laryngopharyngeal reflux and the change of laryngopharyngeal mucosal ultrastructure.

BACKGROUND/AIMS: To establish an animal model of laryngopharyngeal reflux (LPR) and study the effect of LPR on the laryngopharyngeal mucosal ultrastructure. METHODS: Ten Bama minipigs were randomly divided into control group and stent group. Every pig underwent endoscope, and baseline pH was monitored for 4 hours at laryngopharynx and distal esophagus, then specimens from laryngopharyngeal mucosa were biopsied. For the control group, these procedures were repeated on the 14th day. In the stent group, a custom-designed esophageal stent suit was implanted into esophagus, laryngopharyngeal and distal esophageal pH monitoring lasted for 2 hours, then stent suit was removed 3 days later. At last, the same procedures were done as the control group on the 14th day. Specimens were observed under transmission electron microscope to measure the intercellular space and desmosome number. RESULTS: In the control group, there was no laryngopharyngeal reflux on the first day and 14th day. Before the stent was implanted, there was also no laryngopharyngeal reflux in the stent group. In both 2 hours and 14 days after stent implantation, the num-ber of reflux, reflux time, and percentage time of pH < 4.0 were significantly increased (P < 0.05) in the stent group. There was no difference in intercellular space and desmosomes in the control group between baseline and 14th day. In the stent group, intercellular space of laryngopharyngeal mucosa was significantly increased (0.37 µm vs 0.96 µm, P = 0.008), and the number of desmosomes was significantly decreased (20.25 vs 9.5, P = 0.003). CONCLUSIONS: A Bama minipig model of LPR was established by implanting a custom-designed stent suit. LPR might destroy the laryngophar yngeal mucosal barrier.

Morphology of bacterial flora in root canals associated with apical abscesses.

BACKGROUND: Apical abscess is an inflammatory process in the peri-radicular tissues caused by biofilms in the necrotic root canal systems. Therefore, a comprehensive analysis of the bacterial colonization is required for a better understanding of the pathogenesis. This study aimed to investigate the patterns of bacterial infection of root canals of teeth with apical abscesses and to determine whether histological and microbiological findings correlated with clinical conditions. METHODS: Eighteen samples from 18 teeth with apical pathological lesions were analyzed. Nine patients with acute apical abscesses experienced severe pain, and nine patients were asymptomatic with a sinus tract. After extraction, each affected root was divided into two halves. One half was processed for histobacteriologic analysis and examined using light microscopy, and the other half was analyzed using scanning electron microscopy (SEM) to determine the patterns of microbial colonization of the root canals. RESULTS: The appearance of each sample subjected to SEM was consistent with the histobacteriologic findings despite the presence or absence of clinical symptoms. Intraradicular biofilms comprising cocci, rods, and/or filaments of amorphous materials were observed in the apical third of the main root canals in all samples. The bacterial biofilms covering the main root canal walls also penetrated the dentinal tubules to varying depths. The morphologies of biofilms varied, and a unique pattern of intraradicular infection was not identified. CONCLUSION: Intraradicular infections formed complex and variable multispecies biofilms and their presence did not correlate with clinical symptoms.