PAX9 mutations and genetic synergism in familial tooth agenesis.

Familial tooth agenesis (FTA) is one of the most common craniofacial anomalies in humans. Loss-of-function mutations in PAX9 and WNT10A have been known to cause FTA with various expressivity. In this study, we identified five FTA kindreds with novel PAX9 disease-causing mutations: p.(Glu7Lys), p.(Val83Leu), p.(Pro118Ser), p.(Ser197Argfs*23), and c.771+4A>G. Concomitant PAX9 and WNT10A pathogenic variants found in two probands with severe phenotypes suggested an effect of mutational synergism. All overexpressed PAX9s showed proper nuclear localization, excepting the p.(Pro118Ser) mutant. Various missense mutations caused differential loss of PAX9 transcriptional ability. PAX9 overexpression in dental pulp cells upregulated LEF1 and AXIN2 expression, indicating a positive regulatory role for PAX9 in canonical Wnt signaling. Analyzing 176 cases with 63 different mutations, we observed a distinct pattern of tooth agenesis for PAX9-associated FTA: Maxillary teeth are in general more frequently affected than mandibular ones. Along with all second molars, maxillary bicuspids and first molars are mostly involved, while maxillary lateral incisors and mandibular bicuspids are relatively less affected. Genotypically, missense mutations are associated with fewer missing teeth than frameshift and nonsense variants. This study significantly expands the phenotypic and genotypic spectrums of PAX9-associated disorders and reveals a molecular mechanism of genetic synergism underlying FTA variable expressivity.

Characterization of Collapsin Response Mediator Protein 2 in Colorectal Cancer Progression in Subjects with Diabetic Comorbidity.

BACKGROUND: Common demographic risk factors are identified in colorectal cancer (CRC) and type 2 diabetes mellitus (DM), nevertheless, the molecular link and mechanism for CRC-DM comorbidity remain elusive. Dysregulated glycogen synthase kinase-3 beta under metabolic imbalance is suggested to accelerate CRC pathogenesis/progression via regulating collpasin response mediator protein-2 (CRMP2). Accordingly, roles of CRMP2 in CRC and CRC-DM patients were investigated for elucidating the molecular convergence of CRC and DM. METHODS: CRMP2 profile in tumor tissues from CRC and CRC-DM patients was investigated to explore the link between CRC and DM etiology. Meanwhile, molecular mechanism of glucose to regulate CRMP2 profile and CRC characteristics was examined in vitro and in vivo. RESULTS: CRMP2 was significantly lower in tumor lesions and associated with advanced tumor stage in CRC-DM patients. Physiological hyperglycemia suppressed CRMP2 expression/activity and augmented malignant characteristics of CRC cells. Hyperglycemia promotes actin de-polymerization, cytoskeleton flexibility and cell proliferation/metastasis by downregulating CRMP2 profile and thus contributes to CRC disease progression. CONCLUSIONS: This study uncovers molecular evidence to substantiate and elucidate the link between CRC and T2DM, as well as characterizing the roles of CRMP2 in CRC-DM. Accordingly, altered metabolic adaptations are promising targets for anti-diabetic and cancer strategies.

Online discovery of the molecular mechanism for directionally detoxification of Fuzi using real-time extractive electrospray ionization mass spectrometry.

ETHNOPHARMACOLOGICAL RELEVANCE: Aconitum carmichaelii Debeaux, a famous traditional medicinal herb for collapse, rheumatic fever, and painful joints, always raises global concerns about its fatal toxicity from toxic alkaloids when improperly processed. Therefore, it is urgent to clarify the internal molecular mechanism of processing detoxification on Aconitum and develop simple and reliable approaches for clinical application, which is also of great significance to the rational medicinal use of Aconitum. AIM OF THE STUDY: The study aimed at developing a complete molecular mechanism exploration strategy in complex medicinal herb decocting system, clarifying the internal molecular mechanism of processing detoxification on Aconitum, and exploring valid approaches for detoxification. MATERIALS AND METHODS: Aconiti Lateralis Radix Praeparata (Fuzi) was selected as the model for exploring the complex Aconitum detoxification mechanism using an advanced online real-time platform based on extractive electrospray ionization mass spectrometry. The methods realized the sensitive capture of dynamic trace intermediates, accurate qualitative and quantitative analysis, and real-time and long-term monitoring of multi-components with satisfactory accuracy and resistance to complex matrices. RESULTS: Components in the complex Aconitum decocting system were real-timely characterized and fat meat was discovered and verified to directionally detoxify Aconitum while reserving the therapy effect. More importantly, the dynamic detoxification mechanism in the chemically complex Aconitum decoction was molecularly profiled. A novel reaction pathway based on nucleophilic substitution reaction mechanism was proposed. As confirmed by the theoretic calculations at DFT B3LYP/6-31G (d) levels, fatty acids (e.g., palmitic acid) acted as a green, cheap, and high-performance catalyst and promote the decomposition of toxic diester alkaloids to non-toxic and active benzoyl-monoester alkaloids through the discovered mechanism. CONCLUSION: The study exposed a novel detoxification molecular mechanism of Aconitum and provided an effective method for the safe use of Aconitum, which could effectively guide the development of traditional processing technology and compatibility regulation of the toxic herb and had great value to the modernization and standardization development of traditional medicine.

Mechanism of Interleukin-4 Reducing Lipid Deposit by Regulating Hormone-Sensitive Lipase.

Accumulating evidence indicates that inflammation participates in the pathophysiological progress from insulin resistance, obesity, metabolic abnormalities, and type 2 diabetes mellitus. Our previous study reveals that interleukin-4 (IL-4) inhibits adipogenesis and promotes lipolysis to decrease lipid deposits by enhancing the activity of hormone sensitive lipase (HSL). The present study further dissects and characterizes the molecular mechanism of IL-4 in regulating HSL expression and lipolytic activity in the terminal differentiated 3T3-L1 mature adipocytes. Our results showed that IL-4 increased cAMP which then enhanced PKA activity and subsequent phosphorylation of HSL and perilipin. The phosphorylated HSL (p-HSL) translocated from cytoplasm to the surface of lipid droplets and exhibited lipolytic function. After being phosphorylated, p-perilipin also facilitated lipolysis through interacting with p-HSL. The in vitro findings were further verified by in vivo study in which IL-4 exhibited pro-lipolytic activity and enhanced HSL activity. In summary, the net outcome of IL-4 treatment is to reduce lipid storage by promoting lipolysis through enhancing HSL activity via cAMP/PKA pathway, the major route leading to lipolysis.

Massive transfusion in upper gastrointestinal bleeding: a new scoring system.

Background: Massive transfusion in patients with upper gastrointestinal bleeding (UGIB) was not investigated. We developed a new scoring system to predict massive transfusion and to enhance care and early resource mobilization. Methods: Massive transfusion was defined as transfusion with ≥10 units of red blood cells within the first 24 h. Data were extracted from a 10-year, six-hospital database. Logistic regression was applied to derive a risk score for massive transfusion using data from 2006 to 2010, in 24,736 patients (developmental cohort). The score was then validated using data from 2011 to 2015 in 27,449 patients (validation cohort). Area under the receiver operating characteristic (AUROC) curve was performed to assess prediction accuracy. Results: Five characteristics were independently associated (p < .001) with massive transfusion: presence of band-form cells among white blood cells (band form >0), international normalized ratio (INR) >1.5, pulse >100 beats per minute or systolic blood pressure <100 mmHg (shock), haemoglobin <8.0 g/dL and endoscopic therapy. The new scoring system successfully discriminated well between UGIB patients requiring massive transfusion and those who did not in both cohorts (AUROC: 0.831, 95%CI: 0.827-0.836; AUROC: 0.822, 95% CI: 0.817-0.826, respectively). Conclusions: The new scoring system predicts massive transfusion requirement in patients with UGIB well. Key messages Massive transfusion is a life-saving management in massive upper gastrointestinal bleeding. How to identify patients requiring massive transfusion in upper gastrointestinal bleeding is poorly documented. Approximately 3.9% of upper gastrointestinal bleeding patients require massive transfusion. A new scoring system is developed to identify patients requiring massive transfusion with high accuracy.

Comparison of Strain Elastography, Shear Wave Elastography, and Conventional Ultrasound in Diagnosing Thyroid Nodules.

OBJECTIVE: The purpose of this study is to compare the diagnostic performances of strain elastography (SE), shear wave elastography (SWE), and traditional ultrasound (US) features in diagnosing thyroid nodules. SUBJECTS AND METHODS: This study included 185 adult patients with thyroid nodules who underwent conventional gray-scale US, SE, and SWE. SE was scored using a four-pattern elastographic scoring (ES) system. SWE values were presented as mean SWE values and standard derivation using Young's modules. The optimal cutoff values of the mean SWE values for predicting malignancy were determined using receiver operating characteristic (ROC) curve analysis. We used logistic regression models to test elastography as a novel significant predictor for the diagnosis of malignant nodules. The diagnostic performance of elastography parameters was compared with a traditional trained model. RESULTS: Malignant thyroid nodules were stiffer for SE (ES patterns 1 and 2/3 and 4) and mean SWE values (4/17; 51.0 ± 24.4 kPa) than for benign nodules (114/50; 33.1 ± 25.2 kPa) (P < 0.01). In ROC curve analyses, a mean SWE value of 32 kPa was the optimal cutoff point, with diagnostic performance measures of 81% sensitivity, 65% specificity, a 23% positive predictive value (PPV), and 96% negative predictive value (NPV). In multivariate logistic regression, the mean SWE value (≥32 kPa) was an independent predictor for malignancy (odds ratio: 16.8; 95% confidence interval [CI]: 3.6-78.3). However, after the addition of SE and SWE to traditional US features, the C-statistic was not significantly increased compared to the traditional model (0.88, 95% CI: 0.81-0.94 vs. 0.91, 0.85-0.97, P = 0.4). CONCLUSION: In this study, we confirmed SWE as an independent predictor for malignant thyroid nodules. However, in comparing the new extended elastography model to our previous prediction model, the new extended model showed no significant difference in the diagnostic performance.

Role of PARL-PINK1-Parkin pathway in adipocyte differentiation.

OBJECTIVE: Adipogenesis determines the number of adipocytes which is increased when individuals become obese. Mitochondria undergo remarkable morphological and functional changes during adipogenesis. PTEN-induced kinase 1 (PINK1) is pivotal to maintain mitochondrial homeostasis in neural cells. The present study aimed at investigating effects of PINK1 on adipogenesis and energy metabolism. METHODS: Expression of presenilin associated rhomboid-like protein (PARL), PINK1 and Parkin, as well as the interaction among these proteins was temporally examined during adipogenesis. In addition, the alterations of mitochondrial mass and the energy metabolism were also analyzed. RESULTS: Adipogenic process can be dissected into 3 stages according to the participation of PARL-PINK1-Parkin system. (1) When pre-adipocytes are switched to differentiation, f-PINK1 is subjected to PARL cleavage to generate s-PINK1 at the early stage of differentiation (0-4day). Mitochondrial mass is increased for generating ambient energy to meet the demands for cellular remodeling. (2) At the second stage (5-6day), s-PINK1 persistently accumulates in mitochondria and translocates into cytoplasm to mediate Parkin degradation. Mitochondria are fragmented to reduce their mass. (3) At the late stage (7-8day), only residual autophagy activity is remained when excess mitochondria have been eliminated. This mitochondria clearance maintains energy consumption of mature adipocytes at the minimal levels for storing energy. PARL silencing aborts adipogenesis by inhibiting PPARγ expression and the finely-orchestrated events. CONCLUSIONS: Our findings reveal the sequential adipogenic events directed by PARL-PINK1-Parkin system, add more evidence supporting the convergence of pathogenesis leading to neurodegenerative and metabolic diseases, and provide substantial information for developing novel therapeutic strategies by manipulating adipogenesis.

Minimally invasive surgery for esophageal achalasia.

Esophageal achalasia is due to the esophagus of neuromuscular dysfunction caused by esophageal functional disease. Its main feature is the lack of esophageal peristalsis, the lower esophageal sphincter pressure and to reduce the swallow's relaxation response. Lower esophageal muscular dissection is one of the main ways to treat esophageal achalasia. At present, the period of muscular layer under the thoracoscope esophagus dissection is one of the treatment of esophageal achalasia. Combined with our experience in minimally invasive esophageal surgery, to improved incision and operation procedure, and adopts the model of the complete period of muscular layer under the thoracoscope esophagus dissection in the treatment of esophageal achalasia.

Fast Differential Analysis of Propolis Using Surface Desorption Atmospheric Pressure Chemical Ionization Mass Spectrometry.

Mass spectral fingerprints of 24 raw propolis samples, including 23 from China and one from the United States, were directly obtained using surface desorption atmospheric pressure chemical ionization mass spectrometry (SDAPCI-MS) without sample pretreatment. Under the optimized experimental conditions, the most abundant signals were detected in the mass ranges of 70 to 500 m/z and 200 to 350 m/z, respectively. Principal component analyses (PCA) for the two mass ranges showed similarities in that the colors had a significant correlation with the first two PCs; in contrast there was no correlation with the climatic zones from which the samples originated. Analytes such as chrysin, pinocembrin, and quercetin were detected and identified using multiple stage mass spectrometry within 3 min. Therefore, SDAPCI-MS can be used for rapid and reliable high-throughput analysis of propolis.

Computed tomography arterial portography for assessment of portal vein injury after blunt hepatic trauma.

PURPOSE: Intrahepatic portal vein injuries secondary to blunt abdominal trauma are difficult to diagnose and can result in insidious bleeding. We aimed to compare computed tomography arterial portography (CTAP), reperfusion CTAP (rCTAP), and conventional computed tomography (CT) for diagnosing portal vein injuries after blunt hepatic trauma. METHODS: Patients with blunt hepatic trauma, who were eligible for nonoperative management, underwent CTAP, rCTAP, and CT. The number and size of perfusion defects observed using the three methods were compared. RESULTS: A total of 13 patients (seven males/six females) with a mean age of 34.5±14.1 years were included in the study. A total of 36 hepatic segments had perfusion defects on rCTAP and CT, while there were 47 hepatic segments with perfusion defects on CTAP. The size of perfusion defects on CT (239 cm3; interquartile range [IQR]: 129.5, 309.5) and rCTAP (238 cm3; IQR: 129.5, 310.5) were significantly smaller compared with CTAP (291 cm3; IQR: 136, 371) (both, P = 0.002). CONCLUSION: Perfusion defects measured by CTAP were significantly greater than those determined by either rCTAP or CT in cases of blunt hepatic trauma. This finding suggests that CTAP is superior to rCTAP and CT in evaluating portal vein injuries after blunt liver trauma.

A comparison of glycated albumin and glycosylated hemoglobin for the screening of diabetes mellitus in Taiwan.

OBJECTIVE: Glycated albumin (GA) values reflect an average plasma glucose level over approximately 2-4 weeks, and the assay is stable and can be run on serum or plasma. The aim of this study was to determine the universality and the clinical utility of GA in screening for diabetes mellitus. METHODS: Subjects consisted of 2192 male and female residents in Yi-lan County, Northern Taiwan (mean age 60.1 years), of whom 54.2% (n = 1188) had previously been diagnosed and treated for diabetes. Fasting blood samples were obtained to measure HbA1c, plasma glucose, serum GA, insulin, and measures of kidney and liver function. The reference values for these parameters were determined. Data from patients with diabetes and non-diabetic controls were also compared. RESULTS: Mean GA values were 13.8% in controls and 18.1% in diabetic subjects (31.2% higher, p < 0.0001), while mean HbA1c values were 5.6% in controls and 7.2% in diabetic subjects (29.2% higher, p < 0.0001). The 95th percentile values for GA and HbA1c in controls were 16.1% and 6.2%, respectively. Our suggested GA and HbA1c cut-points for prediabetes at the 75th percentile of the normal population would be 14.6% and 5.8%, respectively. For both parameters, values greater than these cut-points provided a reasonable degree of specificity and sensitivity for risk of having diabetes, while a GA value of 16.5% corresponds to an HbA1c level of 6.5%, diagnostic of diabetes. CONCLUSION: These data indicate that GA values can be used as a surrogate parameter for HbA1c in screening for prediabetes and diabetes mellitus.

Video-assisted thoracoscopic pneumonectomy.

BACKGROUND: Lung cancer often requires pneumonectomy. This procedure is challenging and usually performed by thoracotomy, which is traumatic and may involve complications. Video-assisted thoracoscopic surgery (VATS) lobectomy is a recognized procedure that has been accepted by surgeons. There is no standard procedure to perform a pneumonectomy using VATS. The aim of this paper is to share our experiences and to show our technique for performing a pneumonectomy using VATS. METHODS: A 65-year-old man was admitted to the First Affiliated Hospital of Chongqing Medical University. A thoracic computed tomography (CT) scan revealed a 56 mm × 45 mm × 40 mm lesion in the left upper lung lobe. Lesions involving the left lower lung lobe were also identified and the subcarinal and hilar lymph nodes were enlarged. A VATS pneumonectomy was performed. RESULTS: The total surgery time was approximately 90 min, the intraoperative blood loss was 100 mL, the number of resected lymph nodes was 15; and the postoperative hospital stay was 8 days. Follow-up revealed no recurrence or metastasis for 6 months. CONCLUSIONS: Video-assisted thoracoscopic pneumonectomy is a safe and effective treatment procedure.

Vocal cord dysfunction diagnosed by four-dimensional dynamic volume computed tomography in patients with difficult-to-treat asthma: A case series.

Patients with asthma may also have vocal cord dysfunction (VCD), which leads to poor control of the asthma. Once patients are diagnosed with difficult-to-treat asthma with poor control, VCD should be excluded or treated accordingly. The gold standard for diagnosis of VCD is to perform a laryngoscopy. However, this procedure is invasive and may not be suitable for patients with difficult-to-treat asthma. Four-dimensional (4D) dynamic volume computed tomography (CT) is a noninvasive method for quantification of laryngeal movement, and can serve as an alternative for the diagnosis of VCD. Herein, we present a series of five cases with difficult-to-treat asthma patients who were diagnosed with VCD by 4D dynamic volume CT. Clinicians should be alert to the possibility of VCD when poor control is noted in patients with asthma. Early diagnosis by noninvasive 4D dynamic volume CT can decrease excessive doses of inhaled corticosteroids.

[Determination of sodium, magnesium, calcium, lithium and strontium in natural mineral drinking water by microwave plasma torch spectrometer with nebulization sample introduction system].

The microwave plasma torch (MPT) was used as the emission light source. Aqueous samples were introduced with a nebulizer and a desolvation system. A method for the determination of Na, Mg, Ca, Li and Sr in natural mineral drinking water by argon microwave plasma torch spectrometer (ArMPT spectrometer) was established. The effects of microwave power, flow rate of carrier gas and support gas were investigated in detail and these parameters were optimized. Under the optimized condition, the experiments for the determination of Na, Mg, Ca, Li and Sr in 11 kinds of bottled mineral drinking water were carried out by ArMPT spectrometer. The limit-of-detection (LOD) of Na, Mg, Ca, Li and Sr was found to be 4.4, 21, 56, 11 and 84 µg x mL(-1), respectively. Relative standard deviation (n = 6) was in the range of 1.30%-5.45% and standard addition recoveries were in the range of 84.6%-98.5%. MPT spectrometer was simpler, more convenient and of lower cost as compared to ICP unit. MPT spectrometer demonstrated its rapid analysis speed, accuracy, sensitivity and simultaneous multi element analysis ability during the analysis process. The results showed that MPT spectrometer was suitable for metal elements detection for natural mineral drinking water. This approach provides not only one way for resisting the illegal dealings, but also a security for the quality of drinking water. Moreover, the usability of MPT spectrometer in the field of food security; drug safety; clinical diagnostic is promised.

Reactive extractive electrospray ionization tandem mass spectrometry for sensitive detection of tetrabromobisphenol A derivatives.

Sensitive detection of tetrabromobisphenol A (TBBPA) and its derivatives, a group of emerging toxic contaminants, is highly necessitated in environmental investigation. Herein a novel analytical strategy based on reactive extractive electrospray ionization (EESI) tandem mass spectrometry for detection of tetrabromobisphenol A bis(2-hydroxyethyl ether) (TBBPA-BHEE), tetrabromobisphenol A bis(glycidyl ether) (TBBPA-BGE), tetrabromobisphenol A bis(allylether) (TBBPA-BAE), and tetrabromobisphenol S bis(allylether) (TBBPS-BAE) in industrial waste water samples was developed. Active silver cations (Ag(+)), generated by electrospraying a silver nitrate methanol solution (10 mg L(-1)), collides the neutral TBBPA derivatives molecules in the EESI source to form [M+Ag](+) complexes of the analytes under the ambient conditions. Upon collision-induced dissociation (CID), characteristic fragments of the [M+Ag](+) complexes were identified for confident and sensitive detection of the four TBBPA derivatives. Under the optimized experimental conditions, the instrumental limits of detection (LODs) of TBBPA-BHEE, TBBPA-BGE, TBBPA-BAE and TBBPS-BAE were 0.37, 0.050, 0.76, and 4.6 µg L(-1), respectively. The linear ranges extended to 1000 µg L(-1) (R(2)≥0.9919), and the relative standard deviations (RSDs), inter-day variation and intra-day variation were less than 7.8% (n=9), 10.0% (n=5), and 14.8% (n=1 per day for 5 days) for all derivatives. TBBPA derivative manufacturing industrial waste water, river water and tap water samples were fast analyzed with the proposed method. The contents of TBBPA derivatives were various in the collected samples, with the highest 19.9±0.3 µg L(-1) of TBBPA-BAE in the waste water samples.

In vitro suppression of oral squamous cell carcinoma growth by ultrasound-mediated delivery of curcumin microemulsions.

There is increasing interest in using natural products as anticancer agents, as many have antioxidative properties that may help to prevent cellular damage that can lead to cancer. In addition, there is the expectation that many natural products will have low toxicity and few side effects. However, most anticancer and antioxidative agents are hydrophobic, reducing their bioavailability in vivo and making them problematic to deliver. Curcumin provides a good model system for study. In low doses it shows both anticancer and antioxidation effects, whereas in high doses and delivered locally it could be cytotoxic for cancer cells. In this paper, curcumin microemulsions were formed with food-grade chemicals, including soybean lecithin, soybean oil, and Tween 80, a Food and Drug Administration-approved surfactant. The optimized composition formed curcumin microemulsions with a mean size of 40-50 nm, carrying a concentration of curcumin as high as 15 µM. The stability of curcumin microemulsions refrigerated at 5°C over at least 968 days was assessed by size distribution and zeta potential. The effects of low-frequency ultrasound on two oral squamous cell carcinoma cell lines (OSCC-4 and OSCC-25), and the synergy between treatment with curcumin microemulsions and low-frequency sonic stimulation, were tested. Finally, microscopic imaging of the cells confirmed the toxic effects of the curcumin microemulsions, showing damaged and ruptured cells after treatment. Brief exposure to the curcumin-containing microemulsions did have cytotoxic effects, but the addition of ultrasound strongly enhanced those effects, especially on OSCC-25 cells.

Japanese viral encephalitis mimicking stroke with an initial manifestation of hemiplegia.

Japanese encephalitis (JE) is an endemic disease in Taiwan. After the program to vaccinate children against JE was implemented in 1968, the incidence of JE gradually started to decrease, but it is still an important infectious disease here. Neurological manifestations in JE vary highly during the initial stage of the disease. Focal neurological symptoms, such as hemiplegia, are rarely reported. A 46-year-old male with the initial presentation of abrupt hemiplegia and fever developed mental confusion after 1 day. No bacterial pathogen was isolated from the blood or cerebrospinal fluid (CSF). A diagnosis of JE was confirmed based on the presence of JE virus-specific immunoglobulin M in the CSF and serum samples. It is necessary to consider JE when a patient presents with abrupt hemiplegia with fever followed with mental confusion and seizure, especially if the patient comes from a JE-endemic area.