[Clinical study of the Cobb+1 to Cobb fusion strategy for Lenke 5C adolescent idiopathic scoliosis patients with the lower lumbar apex].

Objective: To investigate the indications and surgical outcome of Cobb+1 to Cobb fusion strategy in Lenke 5C adolescent idiopathic scoliosis (AIS) patients with the lower lumbar apex. Methods: The clinical data of Lenke 5C AIS patients treated in Nanjing Drum Tower Hospital from August 2015 to December 2018 were retrospectively analyzed. The patients were followed-up for at least 2 years after surgery and treated with selective Cobb+1 to Cobb fusion strategy. The patients were divided into the normal lumbar apex group (apex location of the main curve was between T12 and L1) and the lower lumbar apex group (apex location of the main curve was below the disc of L1/L2). The occurrence of proximal decompensation in the two groups was compared. In addition, according to whether the patients had proximal decompensation at the last follow-up, the patients in the lower lumbar apex group were further divided into proximal decompensation group and non-decompensation group. The radiographic parameters and Scoliosis Research Society-22 (SRS-22) scores of the two groups were compared. Results: A total of 52 patients (19 cases in the normal lumbar apex group and 33 cases in the lower lumbar apex group), aged (15.3±1.6) years, were followed up for 2-5 (3.2±1.2) years. Six patients (6/19) in the normal lumbar apex group and 5 cases (15.2%) in the lower lumbar apex group showed proximal decompensation during follow-up, and the incidence was significantly higher in the normal lumbar apex group (P=0.034). Within the lower lumbar apex group, the patients with proximal decompensation (n=5) showed similar Risser grade, baseline thoracic Cobb angle, and main Cobb angle as those without proximal decompensation(n=28), and the differences were all not statistically significant (all P>0.05). However, the baseline thoracic/lumbar apical vertebra translation (AVT) ratio was significantly larger in patients with proximal decompensation (0.6±0.2 vs 0.4±0.2, P=0.042), but the postoperative upper instrumented vertebra (UIV) tilt angle was similar (4.5°±2.3° vs 6.2°±3.4°, P=0.312). Conclusion: Cobb+1 to Cobb fusion strategy, selecting UIV at 1 level above upper end vertebra (UEV), could be performed in Lenke 5C patients with the lower lumbar apex location. In addition, UIV could be selected at UEV+1 in patients with small baseline thoracic curve.

Wideband Direct Detection Constraints on Hidden Photon Dark Matter with the QUALIPHIDE Experiment.

We report direction detection constraints on the presence of hidden photon dark matter with masses between 20-30 µeV c^{-2}, using a cryogenic emitter-receiver-amplifier spectroscopy setup designed as the first iteration of QUALIPHIDE (quantum limited photons in the dark experiment). A metallic dish sources conversion photons, from hidden photon kinetic mixing, onto a horn antenna which is coupled to a C band kinetic inductance traveling wave parametric amplifier, providing for near quantum-limited noise performance. We demonstrate a first probing of the kinetic mixing parameter χ to the 10^{-12} level for the majority of hidden photon masses in this region. These results not only represent stringent constraints on new dark matter parameter space, but are also the first demonstrated use of wideband quantum-limited amplification for astroparticle applications.

Nuclear translocation of YAP drives BMI-associated hepatocarcinogenesis in hepatitis B virus infection.

BACKGROUND AND AIMS: Hepatitis B virus (HBV) infection is a major cause of hepatocellular carcinoma (HCC) development and progression. The aim of this study was to mechanistically investigate the involvement of Hippo signalling in HBV surface antigen (HBsAg)-dependent neoplastic transformation. METHODS: Liver tissue and hepatocytes from HBsAg-transgenic mice were examined for the Hippo cascade and proliferative events. Functional experiments in mouse hepatoma cells included knockdown, overexpression, luciferase reporter assays and chromatin immunoprecipitation. Results were validated in HBV-related HCC biopsies. RESULTS: Hepatic expression signatures in HBsAg-transgenic mice correlated with YAP responses, cell cycle control, DNA damage and spindle events. Polyploidy and aneuploidy occurred in HBsAg-transgenic hepatocytes. Suppression and inactivation of MST1/2 led to the loss of YAP phosphorylation and the induction of BMI1 expression in vivo and in vitro. Increased BMI1 directly mediated cell proliferation associated with decreased level of p16INK4a , p19ARF , p53 and Caspase 3 as well as increased Cyclin D1 and γ-H2AX expression. Chromatin immunoprecipitation and the analysis of mutated binding sites in dual-luciferase reporter assays confirmed that the YAP/TEAD4 transcription factor complex bound and activated the Bmi1 promoter. In chronic hepatitis B patients, paired liver biopsies of non-tumour and tumour tissue indicated a correlation between YAP expression and the abundance of BMI1. In a proof-of-concept, treatment of HBsAg-transgenic mice with YAP inhibitor verteporfin directly suppressed the BMI1-related cell cycle. CONCLUSION: HBV-associated proliferative HCC might be related to the HBsAg-YAP-BMI1 axis and offer a potential target for the development of new therapeutic approaches.

[Impact of C2-femoral heads center and sacroiliac fixation on the occurrence of post-operative proximal junctional kyphosis in adults with spinal deformity].

Objective: To analyze the relationship between post-operative proximal junctional kyphosis (PJK) and C2-femoral heads center (C2-FH), pelvic fixation in adult spinal deformity (ASD) patients, so as to provide a reference for the preoperative decision-making and the postoperative intervention timely. Methods: It was a retrospective study that analyzed 34 cases of ASD patients who underwent posterior pedicle screw fixation in Nanjing Drum Tower Hospital between January 2017 and May 2017. ASD patients were divided into two groups according to fixation options: the sacroiliac fixation group and the non-sacroiliac fixation group. The spine-pelvis parameters were evaluated on full-length X ray films of spine at preoperatively, 2 weeks postoperatively and the last follow-up, and the incidence of PJK was recorded. Results: A total of 34 ASD patients (3 males and 31 females) were included in this study, with an average age of (58±6) years. The mean follow-up period was (2.9±1.0) years (2.0-4.0 years). The age, follow-up time and various spine-pelvis parameters showed no significant differences between the two groups (all P>0.05). The immediate postoperative C2-FH in the sacroiliac fixation group was significantly lower than that in the non-sacroiliac fixation group [(-69.46±30.85) mm vs (-31.62±15.31) mm, P<0.001]. The incidence of PJK was as high as 50.0% (8/16) in patients with sacroiliac fixation, but it was only 22% (4/18) in patients without sacroiliac fixation (P=0.016). At the last follow-up, the C2-FH of both groups were both higher than -20 mm (both were approximately -15 mm, P=0.976), indicating that the C2-FH was compensated in both groups. Due to the loss of the distal compensation ability, the incidence of PJK in the sacroiliac fixation group was higher than that in the non-sacroiliac fixation group. Due to the fixation of pelvis and most of lumbar spine, no significant differences were found in lumbar lordosis (LL), pelvic tilt (PT) and sacral slope (SS) at the last follow-up in both groups when compared with those after the operation (all P>0.05); while the thoracic kyphosis (TK) and cervical lordosis (CL) increased significantly (both P<0.05) to compensate C2-FH at the last follow-up. Conclusion: It's demonstrated that C2-FH< -2 cm and pelvic fixation are high risk factors of PJK in ASD patients. It is suggested that both global sagittal balance and pelvic fixation should be considered in decision-making and corrective surgery, rather than only focusing on spine-pelvic parameters.

[Occurrence of rod fracture and salvage methods after primary surgery of the congenital lumbosacral deformity associated with sacral agenesis].

Objective: To analyze the occurrence of rod fracture after surgery for lumbosacral deformity associated sacral agenesis and discuss the relevant salvage methods. Methods: The clinical records of 19 patients who underwent surgical treatment for lumbosacral deformity associated sacral agenesis from January 2001 to January 2018 were retrospectively reviewed, including 11 boys and 8 girls. The average age was (9.6±5.2) years. The outcomes of surgical correction and internal fixation were evaluated by postoperative regular follow-up. We also recorded the time and position of rod fracture occurrence. The Cobb angle, coronal balance and sagittal balance were measured and compared to analyze the corresponding salvage methods and revision outcomes. Results: Three patients encountered rod fracture during follow-up, so the incidence of rod fracture after surgery for lumbosacral deformity associated sacral agenesis was 15.8%(3/19). Based on their own conditions, we formulated the individualized strategy and performed the revision surgery through the posterior-only approach. The most critical step was abundant bone-grafting and fusion in the defected sacroiliac joint. After revision, the scoliotic Cobb angle improved in two patients (91.5° vs 47.5°, 49.0° vs 28.0°) and coronal balance improved in one patient (40.3 mm vs 24.3 mm). No complication reoccurred during follow-up. Conclusion: The rod fracture after surgery for lumbosacral deformity associated sacral agenesis is quite common, which is probably correlated with its unique deformed structure and biomechanical characteristics. The individualized salvage methods and adequate bone-grafting and fusion for the defected sacroiliac joint will guarantee the reconstruction and maintenance of spine balance after revision.

[Clinical study of selection of the upper instrumented vertebra at one level caudal to upper end vertebra in patients with Lenke 5C adolescent idiopathic scoliosis].

Objective: To investigate whether the upper instrumented vertebra (UIV) can be selected at one level caudal to upper end vertebra (UEV) in Lenke type 5C adolescent idiopathic scoliosis (AIS) patients. Methods: Total of 28 Lenke 5C AIS patients who underwent selective posterior fusion in Drum Tower Hospital of Nanjing University Medical School from September 2013 to September 2015 were included. There were 4 males and 24 females, with an age of (15.0±2.0) years, the Risser sign was graded 2-5. The following imaging parameters were measured on standing full spine X-ray before, immediately after the surgery and at the last follow-up: thoracolumbar/lumbar (TL/L) Cobb angle, coronal balance, UIV translation, lower instrumented vertebra (LIV) translation, UIV tilt, LIV tilt, and thoracic apical vertebral translation (T-AVT), lumbar apical vertebral translation (L-AVT). The patients were divided into two groups: decompensation group (n=6) and non-decompensation group (n=22). Radiographic parameters and Scoliosis Research Society (SRS)-22 scores were compared between the two groups. Results: Six cases (21.4%) had proximal decompensation at the last follow-up. There were no significant differences in Risser grade(3.8±1.0 vs 3.6±1.6), baseline thoracic Cobb angle(25.8°±2.2° vs 26.3°±6.4°) and TL/L Cobb angle(43.7°±3.4° vs 45.2°±6.5°) between the two groups (all P>0.05). However, the baseline lumbar/thoracic apical vertebra translation (L-T AVT ratio) was significantly higher in patients with proximal decompensation (6.3±1.3 vs 4.0±2.0, P=0.048). Conclusion: Selecting UIV at one level caudal to UEV, would not increase the incidence of proximal decompensation in Lenke 5C AIS patients with Risser higher than grade 2, the smaller baseline L-T AVT ratio, and with thoracic compensatory curve over 15°, and can obtain satisfactory clinical results.

[Re-discussion on the diagnosis and treatment of cytomegalovirus hepatitis in children].

Cytomegalovirus hepatitis, especially infants with cholestatic liver disease or abnormal liver function, is of great concern to pediatricians. Previously, cytomegalovirus infection was the recognized cause of the disease. Therefore, a comprehensive scientific understanding of cytomegalovirus hepatitis requires an understanding of the basic knowledge of cytomegalovirus infection. Concurrently, it is necessary to standardize the indications of antiviral treatment in combination with the child's age, immune status and other potential diseases to avoid the abuse of antiviral drugs.

The effect of levofloxacin on the lung microbiota of laboratory rats.

Aim: The aim of this study was to investigate the short-term effect of levofloxacin on the microbiota of healthy lungs. Material and methods: Male F344 rats received either no levofloxacin (n = 9), intravenous levofloxacin (n = 12), oral levofloxacin (n = 12), or subcutaneous levofloxacin (n = 14). Rats received a clinically applicable dose (5.56 mg/kg) of levofloxacin via the assigned delivery route once daily for three days. On day four, lung tissue was collected and the lung microbiota composition was investigated using 16S ribosomal RNA gene sequencing. Results: Untreated lungs showed a microbiota dominated by bacteria of the genera Serratia. After treatment with levofloxacin, bacteria of the genus Pantoea dominated the lung microbiota. This was observed for all routes of antibiotic administration, with a significant difference compared to no-antibiotic control group (PERMANOVA: P < 0.001; homogeneity of dispersions: P = 0.656). Conclusion: Our study is the first to demonstrate the effects of levofloxacin therapy on lung microbiota in laboratory rats. Levofloxacin treatment by any route of administration leads to profound changes in the rat lung microbiota, resulting in the predominance of bacteria belonging to the genus Pantoea. Further studies regarding the role of long-term application of broad spectrum antibiotics on induction of lung, allergic and autoimmune diseases are indicated.

Effects of dietary isomaltooligosaccharide and Bacillus spp. supplementation during perinatal period on lactational performance, blood metabolites, and milk composition of sows.

BACKGROUND: In this study, we evaluated the effects of isomaltooligosaccharide (IMO) and Bacillus spp. in perinatal diets on lactational performance, blood metabolites, and milk composition of sows. Multiparous gestating sows (N = 130) with similar body conditions were fed diets without containing IMO (control, CON group), IMO (IMO group), IMO and Bacillus subtilis (IMO + S group), IMO and Bacillus licheniformis (IMO + L group), and IMO and both B. subtilis and B. licheniformis (IMO + S + L group). RESULTS: The average daily feed intake of lactating sows in the IMO + S group (P < 0.05) and the average litter gain and total milk yield in the IMO + S + L group (P < 0.01) were significantly higher than those of the sows in the other treatment groups. In addition, the serum alanine aminotransferase and alkaline phosphatase concentrations in all treatment groups were lower than those in the CON group (P < 0.05). Furthermore, the highest proportion of fat in colostrum (0.05 < P < 0.1) and lowest proportion of lactose in milk were observed in the IMO + L group (P < 0.05). The concentrations of growth hormone and immunoglobulins A and G in milk were higher in sows supplemented with IMO, IMO + S, IMO + L, or IMO + S + L (P < 0.05). CONCLUSION: IMO and Bacillus spp. supplementation during the perinatal period improved the sows' lactational performance by affecting their metabolism and milk quality. Sows in the IMO + S and IMO + S + L treatment groups exhibited the best performance. © 2019 Society of Chemical Industry.

[Investigation on endometrium from menstrual blood as a source of non-invasive tissue].

Objective: To investigate the feasibility of a non-invasive sampling method by collecting menstrual blood and obtaining endometrium for further research in menstruation-related diseases. Methods: On the second day of menstruation, menstrual blood was collected with menstrual cups for 4 hours, and the menstrual endometrium was filtered through a metal screen for weighing, cryopreserved, immunohistochemical staining and cell culture. Results: The collection process was painless and non-invasive. In the control group, the menstrual volume was (9.1±0.7) ml, and the endometrial tissue weight was (91.0±14.7) g. In the endometriosis group, the menstrual volume was (9.6±1.9) ml (P=0.022), and the endometrial tissue weight was (134.7±43.9) g (P=0.057). Endometrial cell culture was successful in all patients and should not be contaminated. The growth curve was a finite cell line type. The expression of cytokeratin 19 and vimentin in menstrual endometrium and cells were positive. Conclusions: By collecting menstrual blood and filtering endometrial tissue, it is an ideal non-invasive sampling method. In combination with advanced experimental technology, menstrual endometrium make further researches of endometriosis, endometrial lesions or other menstruation-related diseases possible.

Effect of gastric fluid aspiration on the lung microbiota of laboratory rats.

AIM OF THE STUDY: The pulmonary microbiota is important for both normal homeostasis and the progression of disease, and may be affected by aspiration of gastric fluid. The aim of this study was to investigate changes in the lung microbiota induced by aspiration of gastric fluid in a laboratory rat model. MATERIAL AND METHODS: Using the intratracheal application method, male rats received aspiration with 0.9% normal saline (n = 11); gastric fluid (n = 24) or sterilized (gamma-irradiated) gastric fluid (n = 12) once-weekly for four weeks. On the fifth week, the animals were sacrificed, and the microbiota of the lung was assessed by 16S ribosomal RNA gene sequencing. RESULTS: Lungs without aspiration and lungs after aspiration with normal saline had similar microbial compositions, dominated by bacteria of the genera Serratia, Ralstonia and Brucella. Evaluation of the microbiota following aspiration of gastric fluid revealed a much different profile that was dominated by bacteria from the genera Romboutsia and Turicibacter and largely independent of sterilization of the gastric fluid. CONCLUSION: In a laboratory rat model, aspiration with gastric fluid caused a substantial shift of the lung microbiota that could be characterized as a shift from Proteobacteria towards Firmicutes, possibly of enteric origin. Bacteria contained in the gastric fluid are not apparently responsible for this change.

[Similar coronal curvature may not represent the same 3-dimensional deformity in adolescent idiopathic scoliosis: a matched-pair study using EOS imaging system].

Objective: To analyze the preoperative axial plane and the surgical outcomes of the Lenke type 1A patients with adolescent idiopathic scoliosis (AIS) whose coronal curve type was matched but thoracic kyphosis (TK) was different. Methods: This study retrospectively reviewed a series of Lenke type 1A female AIS patients who underwent corrective surgery in the Department of Spine Surgery of Nanjing Drum Tower Hospital from May to August 2017. After matched with the Cobb angle of the main thoracic curve, the apical vertebral, the vertebra number included in the curve, the lumber modifier in the Lenke classification and Risser sign, 12 pairs of AIS patients, whose coronal curve was matched but thoracic kyphosis was different, were included in this study. The patients were divided into normal TK group and thoracic hypokyphosis group. EOS whole-body images were taken preoperatively and reconstructed by three-dimensional reconstruction. The whole spine anteroposterior X-ray was taken at 3 weeks after surgery. The radiographic parameters were measured on the preoperative and postoperative two-dimensional X-ray images: coronal Cobb angle, TK, lumbar lordosis (LL), pelvic incidence (PI) and pelvic tilt (PT). The vertebra rotation was obtained on the EOS three-dimensional reconstructed image, and the average vertebral rotation of the major thoracic curve (MTR), the average vertebral rotation of the proximal thoracic curve (PTR) and the average vertebral rotation of the lumbar curve (LR) were calculated. The paired sample t test was used to compare the difference of preoperative and postoperative radiographic parameters between the groups. Results: A total of 24 patients (12 pairs) were included in this study with an average age of (13.7±2.9) years. The preoperative Cobb angle was similar in the two groups (53.8°±10.2° vs 51.0°±11.1°, t=0.27, P=0.81). The average preoperative TK of the normal TK groups was 28.2°±6.1°, while that of the thoracic hypokyphosis group was 11.2°±5.6°(t=7.68, P<0.01). The MTR in the normal TK group was significantly smaller than that in the thoracic hypokyphosis group (10.2° vs 12.7°, t=-3.74, P<0.01), and there was a significant correlation between TK and MTR (r=0.30, P=0.03). As for the lumbar curve rotation, the LR of the normal TK group was significantly smaller than that in the thoracic hypokyphosis group (t=-2.65, P=0.002), but the absolute value of the two groups was similar (t=-0.33, P=0.31). The lumbar Cobb angle correction rate was significantly greater in patients with thoracic and lumbar curve rotating in the same direction than that in the opposite direction (81.1% vs 61.9%, t=4.24, P=0.005). Conclusions: It indicated that when the coronal deformity is matched, the MTR of the patients with thoracic hypokyphosis is significantly larger than that in the patients with normal thoracic kyphosis. The direction of the thoracic and lumbar curve rotation is required to be well concerned in the preoperative surgical planning.

Murine model of oropharyngeal gastric fluid aspiration-A new assessment method for intrapulmonary liquid distribution using digital pixel calculation.

AIM OF THE STUDY: The aim of this study was to investigate a new method for visualization and quantification of intrapulmonary liquid distribution after oropharyngeal gastric fluid aspiration in mice. MATERIAL AND METHODS: Eleven mice received oropharyngeal aspiration with a gastric fluid, India ink, and saline solution. Digital imaging and pixel calculation were used to analyze intrapulmonary fluid distribution selectively. RESULTS: Digital pixel analysis and orophanryngeal aspiration are both safe techniques in mice and deliver reproducible/valid results. Analysis revealed an average aspirate distribution of 86.8% of the total lung area. The proportional amount of the left lung was significantly greater than that of the right lung (P = 0.023). The lobe with the lowest mean distribution was the right lower lobe (79.2% ± 4.4%). CONCLUSION: Digital pixel calculation is a reliable method for quantitative, macroscopic evaluation of fluid distribution in the lung. This method is a useful tool for training purposes and it can be used to ensure interinvestigator reproducibility.

Scutellaria barbata flavonoids alleviate memory deficits and neuronal injuries induced by composited Aß in rats.

BACKGROUND: The aim of the present study was to investigate the effects of Scutellaria barbata flavonoids (SBF) on memory impairment and neuronal injury induced by amyloid beta protein 25-35 in combination with aluminum trichloride (AlCl3) and recombinant human transforming growth factor-ß1 (RHTGF-ß1) (composited Aß) in rats. METHODS: The composited Aß-treated model of Alzheimer's disease (AD)-like memory impairment and neuronal injury was established in male rats by right intracerebroventricular injection of composited Aß, and the effects of SBF were assessed using this rat model. Spatial learning and memory of rats were assessed in the Morris water maze, and neuronal injury was assessed by light and electron microscopy with hematoxylin-eosin or uranyl acetate and lead nitrate-sodium citrate staining, respectively. RESULTS: In the Morris water maze, memory impairment was observed in 94.7% of the composited Aß-treated rats. The composited Aß-treated rats took longer than sham-operated rats to find the hidden platform during position navigation and reversal learning trials. They also spent less time swimming in the target quadrant in the probe trial. Optical and electron microscopic observations showed significant neuropathological changes including neuron loss or pyknosis in hippocampus, typical colliquative necrosis in cerebral cortex, mitochondrial swelling and cristae fragmentation and a large number of lipofuscin deposits in the cytoplasm. Treatment with SBF (35-140 mg/kg) reduced the memory impairment and neuronal injury induced by composited Aß. CONCLUSION: SBF-mediated improvement of composited Aß-induced memory impairment and neuronal injury in rats provides an appropriate rationale for evaluating SBF as a promising agent for treatment of AD.

The role of soluble and insoluble gastric fluid components in the pathogenesis of obliterative bronchiolitis in rat lung allografts.

Repetitive gastric fluid aspirations have been shown to lead to obliterans bronchiolitis (OB), but the component or components of gastric fluid that are responsible are unknown. This study investigates the role of particulates and, separately, soluble material in gastric fluid during the development of OB. Whole gastric fluid (WGF) was collected from male Fischer 344 (F344) rats and separated by centrifugation into particle reduced gastric fluid (PRGF) and particulate components resuspended in normal saline (PNS). Orthotopic left lung transplants from male Wistar-Kyoto rats into F344 rats were performed using a modification of the nonsuture external cuff technique with prolonged cold ischemia. Rats were subjected to weekly aspiration of 0.5 ml/kg of WGF (n = 9), PRGF (n = 10), PNS (n = 9), or normal saline (control, NS; n = 9) for 8 weeks following transplantation. Lung allografts treated with WGF, PRGF, or PNS developed a significantly greater percentage of OB-like lesions compared with the control. No statistical difference was observed when comparing the fibrosis grades or the percentage of OB lesions of WGF, PRGF, and PNS groups, suggesting that both soluble and insoluble components of gastric fluid can promote the development of aspiration-induced OB and fibrosis in lung allografts.

Clearance of bile and trypsin in rat lungs following aspiration of human gastric fluid.

PURPOSE: In the clinical setting, there is no reliable tool for diagnosing gastric aspiration. A potential way of diagnosing gastric fluid aspiration entails bronchoalveolar lavage (BAL) with subsequent examination of the BAL fluid for gastric fluid components that are exogenous to the lungs. The objective of this study was to determine the longevity of the gastric fluid components bile and trypsin in the lung, in order to provide an estimate of the time frame in which assessment of these components in the BAL might effectively be used as a measure of aspiration. MATERIALS AND METHODS: Human gastric fluid (0.5 mg/kg) was infused in the right lung of intubated male Fischer 344 rats (n = 30). Animals were sacrificed at specified times following the experimentally induced aspiration, and bronchoalveolar lavage fluid (BALF) was collected. Bile concentrations were analyzed by an enzyme-linked chromatogenic method, and the concentration of trypsin was quantified using an ELISA. Data were analyzed using non-linear regression and a one-phase decay equation. RESULTS: In this experimental model, the half-life of bile was 9.3 hours (r(2) = 0.81), and the half-life of trypsin was 9.0 hours (r(2) = 0.68). CONCLUSIONS: The half-lives of bile and trypsin in the rodent aspiration model suggest that the ability to detect aspiration may be limited to a few days post-aspiration. If studies using rats are any indication, it may be most effective to collect BAL samples within the first 24 hours of suspected aspiration events in order to detect aspiration.

Mutation analysis of PVRL1 in patients with non-syndromic cleft of the lip and/or palate in Guangdong.

Non-syndromic cleft of the lip and/or palate (NSCLP) is a very common birth defect; the poliovirus receptor-like 1 gene (PVRL1) has been identified as a genetic risk factor for NSCLP in patients from Norway, the Philippines, and South America. Given the considerable variation in allele frequencies across these geographical regions, this study explored the relationship between NSCLP and mutations of PVRL1 in patients from Guangdong, China. We recruited 171 NSCLP patients and 100 volunteers, and divided our samples into 2 groups: a sequencing group and a mass spectrometry group. In the sequencing group, we screened for mutations in exons 2 and 5 of PVRL1 by polymerase chain reaction and direct sequencing in 71 NSCLP patients and 100 volunteers. In the mass spectrometry group, we screened for amino acid mutations in α-spliced transcript codons 112, 131, and 395, and in the ß-spliced transcript codon 1082 using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis in 100 NSCLP patients and 100 volunteers. No mutations were detected in either PVRL1 exons 2 or 5 in the 71 NSCLP patients and 100 volunteers, nor did we find mutations of α-spliced transcript codons 112, 131, 395 and the ß-spliced transcript codon 1082 in any of the 100 NSCLP patients and 100 volunteers. Thus, mutations in exons 2 and 5 of PVRL1, and T334A, A391T, G1183A in the α-spliced transcript, and G1082T in the ß-spliced transcript do not participate in the development of NSCLP in patients from Guangdong.

Epigenetic profiling identifies novel genes for ascending aortic aneurysm formation with bicuspid aortic valves.

BACKGROUND:   Bicuspid aortic valves predispose to ascending aortic aneurysms, but the mechanisms underlying this aortopathy remain incompletely characterized.  We sought to identify epigenetic pathways predisposing to aneurysm formation in bicuspid patients. METHODS:   Ascending aortic aneurysm tissue samples were collected at the time of aortic replacement in subjects with bicuspid and trileaflet aortic valves.  Genome-wide DNA methylation status was determined on DNA from tissue using the Illumina 450K methylation chip, and gene expression was profiled on the same samples using Illumina Whole-Genome DASL arrays.  Gene methylation and expression were compared between bicuspid and trileaflet individuals using an unadjusted Wilcoxon rank sum test.   RESULTS:   Twenty-seven probes in 9 genes showed significant differential methylation and expression (P<5.5x10-4).  The top gene was protein tyrosine phosphatase, non-receptor type 22 (PTPN22), which was hypermethylated (delta beta range: +15.4 to +16.0%) and underexpressed (log 2 gene expression intensity: bicuspid 5.1 vs. trileaflet 7.9, P=2x10-5) in bicuspid patients, as compared to tricuspid patients.  Numerous genes involved in cardiovascular development were also differentially methylated, but not differentially expressed, including ACTA2 (4 probes, delta beta range:  -10.0 to -22.9%), which when mutated causes the syndrome of familial thoracic aortic aneurysms and dissections CONCLUSIONS:   Using an integrated, unbiased genomic approach, we have identified novel genes associated with ascending aortic aneurysms in patients with bicuspid aortic valves, modulated through epigenetic mechanisms.  The top gene was PTPN22, which is involved in T-cell receptor signaling and associated with various immune disorders.  These differences highlight novel potential mechanisms of aneurysm development in the bicuspid population.

DCT protects human melanocytic cells from UVR and ROS damage and increases cell viability.

Dopachrome tautomerase (DCT) is involved in the formation of the photoprotective skin pigment eumelanin and has also been shown to have a role in response to apoptotic stimuli and oxidative stress. The effect of DCT on UVR DNA damage responses and survival pathways in human melanocytic cells was examined by knockdown experiments using melanoma cells, neonatal foreskin melanoblasts (MB) in monoculture and in co-culture with human keratinocytes. MB cell strains genotyped as either MC1R WT or MC1R RHC homozygotes, which are known to be deficient in DCT, were transduced with lentivirus vectors for either DCT knockdown or overexpression. We found melanoma cell survival was reduced by DCT depletion and by UVR over time. UVR-induced p53 and pp53-Ser15 levels were reduced with DCT depletion. Knockdown of DCT in MC1R WT and MC1R RHC MB cells reduced their survival after UVR exposure, whereas increased DCT protein levels enhanced survival. DCT depletion reduced p53 and pp53-Ser15 levels in WM266-4 melanoma and MC1R WT MB cells, while MC1R RHC MB cells displayed variable levels. Both MC1R WT and RHC genotypes of MB cells were responsive to UVR at 3 h with increases in both p53 and pp53-Ser15 proteins. MC1R WT MB cell strains in coculture with keratinocytes have an increased cell survival after UVR exposure when compared to those in monoculture, a protective effect which appears to be conferred by the keratinocytes.

Preprocedural ECG-gated computed tomography for prevention of complications during lead extraction.

BACKGROUND: Preprocedural multidetector computed tomography (MDCT) may identify patients at risk for mechanical complications during lead extraction. METHODS: To describe the use and feasibility of computed tomography scanning for preprocedural planning of lead extraction, we conducted a retrospective study of high-risk patients, who underwent electrocardiogram (ECG)-gated MDCT before planned lead extraction between January 1, 2012, and March 30, 2013. RESULTS: Among 30 patients the mean age was 63 ± 15 years, 60% were male, and 20% had prior sternotomy. Most devices were left sided (93%) and 24 had implantable defibrillators (80%). Indications for extraction included lead malfunction (n = 15; 50%), class I lead advisories (n = 11; 37%), and infection (n = 10; 33%). Overall, there were 65 leads extracted (mean 2.1 leads per patient). One extraction procedure was deferred due to MDCT evidence of significant myocardial perforation with the lead tip > 1 cm beyond the epicardium (n = 1, 3%). MDCT suggestion of lead adherence to central venous structures (n = 13, 43%) was associated with significantly longer laser times (88 ± 71 seconds vs 30 ± 37 seconds, P = 0.02) and larger sheath size (14.9 ± 1.3 vs 13.5 ± 1.2 French, P = 0.02). MDCT evidence of central venous occlusion or stenosis was not associated with increased laser times. Excluding the patient with MDCT evidence of significant perforation, clinical success was achieved in all patients (n = 29/29). CONCLUSIONS: ECG-gated MDCT scanning before lead extraction may facilitate the identification of significant perforation and patients at high risk for mechanical complication.