3D genomic mapping reveals multifocality of human pancreatic precancers.

Pancreatic intraepithelial neoplasias (PanINs) are the most common precursors of pancreatic cancer, but their small size and inaccessibility in humans make them challenging to study1. Critically, the number, dimensions and connectivity of human PanINs remain largely unknown, precluding important insights into early cancer development. Here, we provide a microanatomical survey of human PanINs by analysing 46 large samples of grossly normal human pancreas with a machine-learning pipeline for quantitative 3D histological reconstruction at single-cell resolution. To elucidate genetic relationships between and within PanINs, we developed a workflow in which 3D modelling guides multi-region microdissection and targeted and whole-exome sequencing. From these samples, we calculated a mean burden of 13 PanINs per cm3 and extrapolated that the normal intact adult pancreas harbours hundreds of PanINs, almost all with oncogenic KRAS hotspot mutations. We found that most PanINs originate as independent clones with distinct somatic mutation profiles. Some spatially continuous PanINs were found to contain multiple KRAS mutations; computational and in situ analyses demonstrated that different KRAS mutations localize to distinct cell subpopulations within these neoplasms, indicating their polyclonal origins. The extensive multifocality and genetic heterogeneity of PanINs raises important questions about mechanisms that drive precancer initiation and confer differential progression risk in the human pancreas. This detailed 3D genomic mapping of molecular alterations in human PanINs provides an empirical foundation for early detection and rational interception of pancreatic cancer.

Three-dimensional genomic mapping of human pancreatic tissue reveals striking multifocality and genetic heterogeneity in precancerous lesions.

Pancreatic intraepithelial neoplasia (PanIN) is a precursor to pancreatic cancer and represents a critical opportunity for cancer interception. However, the number, size, shape, and connectivity of PanINs in human pancreatic tissue samples are largely unknown. In this study, we quantitatively assessed human PanINs using CODA, a novel machine-learning pipeline for 3D image analysis that generates quantifiable models of large pieces of human pancreas with single-cell resolution. Using a cohort of 38 large slabs of grossly normal human pancreas from surgical resection specimens, we identified striking multifocality of PanINs, with a mean burden of 13 spatially separate PanINs per cm3 of sampled tissue. Extrapolating this burden to the entire pancreas suggested a median of approximately 1000 PanINs in an entire pancreas. In order to better understand the clonal relationships within and between PanINs, we developed a pipeline for CODA-guided multi-region genomic analysis of PanINs, including targeted and whole exome sequencing. Multi-region assessment of 37 PanINs from eight additional human pancreatic tissue slabs revealed that almost all PanINs contained hotspot mutations in the oncogene KRAS, but no gene other than KRAS was altered in more than 20% of the analyzed PanINs. PanINs contained a mean of 13 somatic mutations per region when analyzed by whole exome sequencing. The majority of analyzed PanINs originated from independent clonal events, with distinct somatic mutation profiles between PanINs in the same tissue slab. A subset of the analyzed PanINs contained multiple KRAS mutations, suggesting a polyclonal origin even in PanINs that are contiguous by rigorous 3D assessment. This study leverages a novel 3D genomic mapping approach to describe, for the first time, the spatial and genetic multifocality of human PanINs, providing important insights into the initiation and progression of pancreatic neoplasia.

Gastroprotective Effects of Fermented Lotus Root against Ethanol/HCl-Induced Gastric Mucosal Acute Toxicity in Rats.

Gastric ulcers are a common gastrointestinal disease across the globe. Alcohol consumption is the primary cause of gastric carcinogenesis and progression. We investigated the gastroprotective effects of fermented lotus root (FL) against ethanol (EtOH)/HCl-induced gastric ulcers in a rat model and the conceivable underlying mechanisms involved. Rats received different doses of FL (50, 100, and 200 mg/kg) or ranitidine (positive control, 30 mg/kg) via oral gavage daily for 14 days. One hour after the last oral administration of FL, the EtOH/HCl mixture was orally intubated to induce gastric damage. Oral administration of FL significantly alleviated the gastric lesions. Moreover, FL also elevated the amounts of nitric oxide and the antioxidant enzyme activities of superoxide dismutase, glutathione peroxidase, and catalase in the stomach. To verify the gastric mucosal defense mechanism, inflammation-related genes were measured. Our results revealed that FL effectively inhibited gastric mucosal damage via downregulation of the nuclear factor-kappaB (NF-κB) response in the stomach. The administration of FL significantly lowered the gastric mRNA expression of inflammation-related genes, including NF-κb1, tumor necrosis factor-α, interferon γ, and prostaglandin-endoperoxide synthase 2, compared with the gastric ulcer control group. In addition, the NF-κB signaling pathway-related protein markers inhibitor of κB (IκB)-α, IκB kinase, and NF-κB were significantly reduced in the FL groups. Taken together, these data suggest that FL administration may have potential as an alternative treatment for gastric ulcers due to its antioxidant and anti-inflammatory effects and its ability to promote the recovery of gastric mucosa.

Ameliorative effects of black ginseng on nonalcoholic fatty liver disease in free fatty acid-induced HepG2 cells and high-fat/high-fructose diet-fed mice.

BACKGROUND: Black ginseng (BG) is a type of Korean ginseng prepared by steaming and drying raw ginseng to improve the saponin content. This study examined the effects of BG on nonalcoholic fatty liver disease (NAFLD) in HepG2 cells and diet-induced obese mice. METHODS: HepG2 cells were treated with free fatty acids to induce lipid accumulation before supplementation with BG. NAFLD-induced mice were fed different doses (0.5%, 1%, and 2%) of BG for 8 weeks. RESULTS: BG significantly reduced lipid accumulation and expression of lipogenic genes, peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein alpha, sterol regulatory element-binding protein-1c, and fatty acid synthase in HepG2 cells, and the livers of mice fed a 45% high-fat diet with 10% fructose in the drinking water (HFHF diet). BG supplementation caused a significant reduction in levels of aspartate aminotransferase and alanine aminotransferase, while antioxidant enzymes activities were significantly increased in 45% high-fat diet with 10% fructose in the drinking water diet-fed mice. Expression of proliferator-activated receptor alpha and carnitine palmitoyltransferase I were upregulated at the transcription and translation levels in both HepG2 cells and diet-induced obese mice. Furthermore, BG-induced phosphorylation of AMP-activated protein kinase and acetyl CoA carboxylase in both models, suggesting its role in AMP-activated protein kinase activation and the acetyl CoA carboxylase signaling pathway. CONCLUSION: Our results indicate that BG may be a potential therapeutic agent for the prevention of NAFLD.

Phellinus baumii enhances the immune response in cyclophosphamide-induced immunosuppressed mice.

Phellinus species is a mushroom used as traditional medicine in Eastern Asia. Research on Phellinus baumii (PB) is relatively limited; however, it has been reported to have antioxidant, DNA damage-protecting, immunostimulating, and antidiabetic activities. In our previous study on anti-inflammatory properties in lipopolysaccharide-stimulated RAW 264.7 cells and the various bioactive components of PB, we propose that PB could exert immune enhancing effects. Therefore, our current study aimed to investigate the immune-enhancing effect on immunosuppressed mice. Different concentrations of PB extract (0, 50, 100, 200, and 400 mg/kg body weight) were given to mice via oral gavage for 6 weeks accompanied by intraperitoneal cyclophosphamide administration to induce immunosuppression. A bone marrow micronucleus test was performed in mice to screen for potential genotoxic compounds. Splenocyte viability and proliferation, splenic and peritoneal natural killer cell activities, and hematological markers were then measured. Cytokines in the spleen and serum, as well as splenic mRNA levels of nuclear factor-κB; interferon-γ; tumor necrosis factor-α; and interleukin (IL)-1ß, IL-6, and IL-12, were determined in mice. As a result, PB ameliorated T- and B-lymphocyte proliferation, splenic and peritoneal NK cell activities, bone marrow cells, hematological markers, cytokine levels, and T-lymphocyte numbers. Moreover, serum and spleen cytokine levels and mRNA expression were elevated in the PB groups compared to controls. Our results suggest that the PB extract can be used as a potent immunomodulator under immunosuppressive conditions. Thus, PB may be used as a potent biofunctional and pharmaceutical material to potentially enhance human immunity.

Comparing effectiveness of polydeoxyribonucleotide injection and corticosteroid injection in plantar fasciitis treatment: A prospective randomized clinical study.

BACKGROUND: This study aimed to compare the efficacy and safety of polydeoxyribonucleotide (PDRN) injection and corticosteroid injection for plantar fasciitis. METHODS: This study included 44 patients with plantar fasciitis, randomly allocated to the PDRN and corticosteroid groups. Evaluation using the visual analogue scale (VAS) pain score and Manchester-Oxford foot questionnaire (MOXFQ) was conducted at baseline, 1, 2, 6weeks and 6months. The thickness and echogenicity of the plantar fascia in ultrasonography and complications were recorded. RESULTS: Corticosteroid injection elicited more pain relief than did PDRN injection at 2 (p=0.010) and 6weeks (p=0.016); however, it showed no superiority at 6months (p=0.523). MOXFQ showed similar outcomes. The thickness and echogenicity did not differ between groups and no complications were reported in either group. CONCLUSIONS: We demonstrated that PDRN injection could be an effective and safe option for plantar fasciitis and was comparable to corticosteroid injection after 6months follow up. LEVEL OF EVIDENCE: II, comparative study.

Optimal Screw Fixation of Syndesmosis Using a Targeting Drill Guide: A Technical Note.

Optimal screw fixation of the syndesmosis is difficult. We introduce a novel technique using a targeting drill guide for centroidal screw fixation of a syndesmosis to ensure a reproducible and optimal screw trajectory for syndesmosis fixation. By using a drill guide for anterior cruciate ligament surgery and intraoperative fluoroscopy, syndesmosis fixation enables reproducible fixation along an individual centroidal axis.

Lonicera caerulea Extract Attenuates Non-Alcoholic Fatty Liver Disease in Free Fatty Acid-Induced HepG2 Hepatocytes and in High Fat Diet-Fed Mice.

Honeyberry (Lonicera caerulea) has been used for medicinal purposes for thousands of years. Its predominant anthocyanin, cyanidin-3-O-glucoside (C3G), possesses antioxidant and many other potent biological activities. We aimed to investigate the effects of honeyberry extract (HBE) supplementation on HepG2 cellular steatosis induced by free fatty acids (FFA) and in diet-induced obese mice. HepG2 cells were incubated with 1 mM FFA to induce lipid accumulation with or without HBE. Obesity in mice was induced by a 45% high fat diet (HFD) for 6 weeks and subsequent supplementation of 0.5% HBE (LH) and 1% HBE (MH) for 6 weeks. HBE suppressed fatty acid synthesis and ameliorated lipid accumulation in HepG2 cells induced by FFA. Moreover, HBE also decreased lipid accumulation in the liver in the supplemented HBE group (LH, 0.5% or MH, 1%) compared with the control group. The expressions of adipogenic genes involved in hepatic lipid metabolism of sterol regulatory element-binding protein-1 (SREBP-1c), CCAAT/enhancer-binding protein alpha (C/EBPα), peroxisome proliferator-activated receptor gamma (PPARγ), and fatty acid synthase (FAS) were decreased both in the HepG2 cells and in the livers of HBE-supplemented mice. In addition, HBE increased mRNA and protein levels of carnitine palmitoyltransferase (CPT-1) and peroxisome proliferator-activated receptor α (PPARα), which are involved in fatty acid oxidation. Furthermore, HBE treatment increased the phosphorylation of AMP-activated protein kinase (AMPK) and Acetyl CoA Carboxylase (ACC). Honeyberry effectively reduced triglyceride accumulation through down-regulation of hepatic lipid metabolic gene expression and up-regulation of the activation of AMPK and ACC signaling in both the HepG2 cells as well as in livers of diet-induced obese mice. These results suggest that HBE may actively ameliorate non-alcoholic fatty liver disease.

Is the Bohler angle reliable for operative reduction of calcaneus fracture?

BACKGROUD: To evaluate reliability of Bohler angle in doing operative reduction of fractured calcaneus, and compare its value over other image lines of hindfoot when surgeons determine whether the injured calcaneus is reduced close to pre-injured state. METHODS: Eighteen of postoperative lateral radiographs were obtained and edited with two versions; one of which anterior of calcaneus erased not to check Bohler angle, the other of which peri-calcaneus structure such as talus erased only to check Bohler angle. Four orthopaedic surgeons were asked to determine the two sets of images whether the injured calcaneus is reduced close to pre-injured state in independent two sessions. Intra-, interobserver reliability, diagnosis test including specificity, sensitivity, positive and negative predictive values were assessed. RESULTS: Intra- and interobserver reliability, Kappa value, ranged from 0.110 to 0.723 regardless the kind of the test. Sensitivity and specificity of both two tests were not significantly different in both trials, either. Positive and negative predictive values also showed similar pattern. The probability that observer determine the specific image as an acceptable reduction of calcaneus-less than 5 degrees of difference of the Bohler angle between post-reduction and pre-injured state-of two tests, was not significantly different in both trials (p = 0.40, 0.24, respectively). CONCLUSIONS: Bohler angle is known as one of the most objective markers for calcaneus fracture, but was not accurate as a sole reference in intra-operative reduction in this study. Therefore, surgeons should take into account the other radiographic features in surgery. LEVELS OF EVIDENCE: IV, case series.

Protective effect of bovine milk against HCl and ethanol-induced gastric ulcer in mice.

The purpose of this study was to investigate the gastroprotective effects of bovine milk on an acidified ethanol (HCl-ethanol) mixture that induced gastric ulcers in a mouse model. Mice received different doses of commercial fresh bovine milk (5, 10, and 20 mL/kg of body weight) by oral gavage once a day for 14 d. One hour after the last oral administration of bovine milk, the HCl-ethanol mixture was orally intubated to provoke severe gastric damage. Our results showed that pretreatment with bovine milk significantly suppressed the formation of gastric mucosa lesions. Pretreatment lowered gastric myeloperoxidase and increased gastric mucus contents and antioxidant enzymes catalase and superoxide dismutase. Administration of bovine milk increased nitrate/nitrite levels and decreased the malondialdehyde levels and the expression of proinflammatory genes, including transcription factor nuclear factor-κB, cyclooxygenase-2, and inducible nitric oxide synthase in the stomach of mice. These results suggest that bovine milk can prevent the development of gastric ulcer caused by acid and alcohol in mice.

Is osteoporosis a risk factor for ankle fracture?: Comparison of bone mineral density between ankle fracture and control groups.

OBJECTIVES: There remains controversy over osteoporotic feature of the ankle. Therefore, we investigated the possibility of the existence of a relationship between axial bone mineral density (BMD) in patients with ankle fracture group with that of the normal population in Korea under control of other confounding factors such as body mass index (BMI). METHODS: This study retrospectively reviewed medical records of patients who were treated in our institution from 2005 to 2015. A comparative analysis was carried out between 116 patients with ankle fracture (ankle fracture group) and 113 patients admitted with other orthopedic reasons (control group). Sex, age, energy level of trauma, and BMI were analyzed as variables affecting axial BMD. RESULTS: Age and sex of ankle fracture group were not different from them of control group (P = 0.968 and P = 0.870, respectively). BMI of ankle fracture group was higher than that of control (P = 0.029). The other variables showed no differences between the 2 groups. The energy level of trauma in ankle fracture group was related to only BMI (P = 0.013). CONCLUSIONS: Axial BMD of ankle fracture patients showed no difference from that of a control group in Korean population. The occurrence of ankle fracture is affected by only BMI rather than axial BMD. Evaluation of osteoporosis for patients with ankle fracture should be considered separately.

Hawthorn Fruit Extract Elevates Expression of Nrf2/HO-1 and Improves Lipid Profiles in Ovariectomized Rats.

The purpose of this study was to investigate the effects of hawthorn (Crataegus pinnatifida Bunge) extract on the lipid profiles and antioxidant properties in ovariectomized (OVX) rats. After ovariectomy, the rats were randomly divided into four groups: the non-OVX control (Sham), the OVX-control (OVX), the OVX + 100 mg/kg b.w. of hawthorn extract (OL), and the OVX + 200 mg/kg b.w. of hawthorn extract (OH). The final body weights of the OVX group were significantly increased, but the increment was significantly decreased in hawthorn groups (p < 0.05). The serum total and low-density lipoprotein (LDL) cholesterol levels were significantly elevated in the OVX group, whereas the hawthorn groups showed a significant decrease in these levels (p < 0.05). The hepatic triglyceride (TG) and malondialdehyde (MDA) levels were significantly reduced in the hawthorn groups compared with the OVX group (p < 0.05). The mRNA expression of nuclear factor erythroid 2-related factor (Nrf2), heme oxygenase-1 (HO-1), and glutathione peroxidase (GPx) were significantly decreased in the OVX group, whereas the hawthorn groups exhibited a significant increase in expression (p < 0.05). The protein expressions of Nrf2, HO-1, and GPx were lower in the OVX group than the Sham group (p < 0.05). The oral administration of hawthorn extract reversed the suppression of protein levels. These results suggest that hawthorn extract could have protective effects in OVX rats by improving lipid profiles, decreasing oxidative stress, and improving the antioxidant defense system.

Magnetic Resonance Imaging Findings in Small Patella Syndrome.

Small patella syndrome (SPS) is characterized by aplasia or hypoplasia of the patella and pelvic girdle abnormalities, including bilateral absence or delayed ossification of the ischiopubic junction and infra-acetabular axe-cut notches. Here, we report a case of SPS in a 26-year-old female. Magnetic resonance image (MRI) showed a small patella with thick eccentric non-ossified patellar cartilage and femoral trochlear dysplasia with hypoplastic patellar undersurface. To our knowledge, this is the first report of MRI findings in SPS. MRI findings could be clinically relevant because elongation of the medial patellofemoral ligament and trochlear dysplasia with eccentric non-ossified patellar cartilage might lead to patellofemoral maltracking with an osteochondral lesion or acute dislocation or an extensor mechanism injury. Though the patient presented in this case report only had a gastrocnemius injury at the origin site, physicians should carefully examine abnormalities with MRI when an SPS patient has a trauma to the knee.

CT analysis of retropharyngeal abnormality in Kawasaki disease.

OBJECTIVE: To retrospectively compare the imaging characteristics of retropharyngeal density and associated findings for Kawasaki disease with those for non-Kawasaki disease, and identify the distinguishing features which aid the CT diagnosis of Kawasaki disease with retropharyngeal low density. MATERIALS AND METHODS: Among the enhanced neck CT performed in children less than 8-years old with clinical presentation of fever and cervical lymphadenopathy over a 6-year period, only cases with retropharyngeal low density (RLD) were included in this study. The 56 cases of RLD were divided into two groups; group A included cases diagnosed as Kawasaki disease (n = 34) and group B included cases diagnosed as non-Kawasaki disease (n = 22). We evaluated the CT features including the thickness of RLD and its extent into the deep neck spaces, as well as soft tissue change in the adjacent structure. We also scored the extent of RLD into the deep neck spaces and the soft tissue changes in the adjacent structure. RESULTS: The thickness of RLD was greater in group A than in group B (group A, 6.0 ± 2.1; group B, 4.6 ± 1.5, p = 0.01). The score of the RLD extent into the deep neck spaces was significantly greater in group A than in group B (group A, 2.3 ± 1.3; group B, 0.8 ± 1.0, p < 0.01). Also, the score of the adjacent soft tissue changes was greater in group A than in group B (group A, 2.0 ± 1.1; group B, 1.0 ± 1.0, p < 0.01). CONCLUSION: If children present with fever and cervical lymphadenopathy that display retropharyngeal low density with extension into more deep neck spaces as well as changes in more adjacent soft tissue, the possibility of Kawasaki disease should be considered.

Factors leading to decreased rates of deep vein thrombosis and pulmonary embolism after total knee arthroplasty.

We evaluated a total of 473 knees (264 patients) to determine the prevalence of deep vein thrombosis and pulmonary embolism (PE) in those who were not given thromboprophylaxis after total knee arthroplasty (TKA). Deep vein thrombosis was diagnosed by roentgenographic venography, and PE was diagnosed by perfusion lung scanning. In patients with bilateral TKA, 47 (11%) of 418 venograms showed positive findings for thrombi, whereas in patients with unilateral TKA, 11 (20%) of 55 venograms showed positive findings for thrombi (P = .758). No patient had symptoms of PE, and findings for the perfusion lung scans were negative in all patients. We neither treat our patients for deep vein thrombosis and PE prophylactically nor therapeutically unless patient has a symptomatic PE.

Sonographic measurements of normal gallbladder sizes in children.

PURPOSE: Our goal was to establish the range of sonographic measurements of normal gallbladders in children. METHODS: Six hundred ten children aged 0-16 years (male:female ratio, 1.5:1) with normal clinical and laboratory findings were included in this study. The sonographic parameters were the length, width, and calculated volume of the gallbladder, and the clinical parameters were the age, height, weight, and body surface area of the children. Statistical significance was determined through correlation and regression analyses. RESULTS: The length of the gallbladder showed significant positive correlations with age (r = 0.65), height (r = 0.67), weight (r = 0.63), and body surface area (r = 0.65; p < 0.01). The calculated volume of the gallbladder also showed moderate correlations with age (r = 0.53), height (r = 0.55), weight (r = 0.61), and body surface area (r = 0.57; p < 0.01). The gallbladder width showed modest but significant correlations with age (r = 0.48), height (r = 0.53), weight (r = 0.53), and body surface area (r = 0.55; p < 0.01). The highest correlation coefficients were found between the gallbladder length and subject age (r = 0.65; p < 0.01) and between the gallbladder length and subject height (r = 0.67; p < 0.01). For all correlations, statistical significance remained after regression analysis (p < 0.01). CONCLUSIONS: Values for the size of the normal pediatric gallbladder are defined and will be helpful in the diagnosis of gallbladder abnormalities.