Phenotype and genetic characteristics in 20 Chinese patients with 46,XY disorders of sex development.

PURPOSE: 46,XY disorders of sex development (DSD) is the most complicated and common type of DSD. To date, more than 30 genes have been identified associated with 46,XY DSD. However, the mutation spectrum of 46,XY DSD is incomplete owing to the high genetic and clinical heterogeneity. This study aims to provide clinical and mutational characteristics of 18 Chinese patients with 46,XY DSD. METHODS: A total of 20 unrelated individuals with 46,XY DSD were recruited. Whole-exome sequencing (WES) or custom-panel sequencing combined Sanger sequencing were performed to detect the pathogenic mutations. The pathogenicity of the variant was assessed according to the American College of Medical Genetics and Genomics (ACMG) guidance and technical standards recommended by the ACMG and the Clinical Genome Resource (ClinGen). RESULTS: Six patients harbored NR5A1 mutations; two patients harbored NR0B1 mutations; six patients harbored SRD5A2 mutations; six patients harbored AR mutations. Six novel genetic variants were identified involved in three genes (NR5A1, NR0B1, and AR). CONCLUSION: We determined the genetic etiology for all enrolled patients. Our study expanded the mutation spectrum of 46,XY DSD and provided diagnostic evidence for patients with the same mutation in the future.

Angiographic features of pulmonary arteriovenous malformations: a new imaging classification system, BRAND, to guide embolisation.

AIM: To introduce a new classification system for pulmonary arteriovenous malformations (PAVMs) based on the BRanches, ANgles and Diameter of the feeding artery (BRAND), which is intended provide a better understanding of the vascular structure of PAVMs and facilitate the selection of therapeutic strategies for embolisation. MATERIALS AND METHODS: Twenty-one patients with PAVMs from 2005 to 2020 were reviewed retrospectively. According to angiographic morphology, classified the PAVMs using the BRAND classification. RESULTS: A total of 41 PAVMs were treated with transcatheter embolisation. Based on the BRAND classification, there were four branches (9.8%), four acute angles (9.8%), and 32 occlusion sites >4 mm in diameter (78%). A total of 30 PAVMs (73.2%) were treated with occluders and plugs while the rest were embolised with coils (26.8%). Immediate technical success was achieved in all patients. In feeding arteries with acute angles, balloon-assisted tracking and trans-septal puncture techniques were used to facilitate the advancement of delivery systems. All origins of branches were preserved. Mean SaO2 increased significantly from 79.5 ± 7.5% before embolisation to 92 ± 5.3% after embolisation (p<0.05). During a median follow-up of 5.4 years, two patients (9.5%) developed new PAVMs and required a second procedure. One patient (4.8%) underwent repeat embolisation due to the reperfusion of previously embolised PAVMs with coils. CONCLUSIONS: BRAND classification is well suited to address the challenging feeding arteries of PAVMs and allow precise placement of the delivery system, leading to minimal procedural risk and high success rates.

Antithrombotic management of patients with atrial fibrillation-Dutch anticoagulant initiatives anno 2020.

In recent years, as more and more experience has been gained with prescribing direct oral anticoagulants (DOACs), new research initiatives have emerged in the Netherlands to improve the safety and appropriateness of DOAC treatment for stroke prevention in patients with atrial fibrillation (AF). These initiatives address several contemporary unresolved issues, such as inappropriate dosing, non-adherence and the long-term management of DOAC treatment. Dutch initiatives have also contributed to the development and improvement of risk prediction models. Although fewer bleeding complications (notably intracranial bleeding) are in general seen with DOACs in comparison with vitamin K antagonists, to successfully identify patients with high bleeding risk and to tailor anticoagulant treatment accordingly to mitigate this increased bleeding risk, is one of the research aims of recent and future years. This review highlights contributions from the Netherlands that aim to address these unresolved issues regarding the anticoagulant management in AF in daily practice, and provides a narrative overview of contemporary stroke and bleeding risk assessment strategies.

[Research Progress and Prospect of Facial Reconstruction in Forensic Science].

ABSTRACT: Facial reconstruction is a way to recover facial morphology by restoring soft tissues based on unidentified skulls using the knowledge of anatomy, anthropology, aesthetics, and computer science. It is applied in forensic science, oral plastic surgery and archeology, and especially plays an important role in the identification of the origin of the unknown corpses in forensic science. Facial reconstruction is the supplementary means of identification when other approaches （such as DNA comparison, imaging matching, dental records comparison, etc.） cannot identify individual identity. Facial soft tissue thickness （FSTT） is the basis of facial reconstruction and with the development of imaging and computer science, the techniques for measuring FSTT are improving rapidly and many related researches have appeared. This paper summarizes the application of facial reconstruction in forensic science, the accuracy of different methods and the research progress of this field to provide reference to this field.

Construction of Age Estimation Model of 18-Year-Olds in the Northern Chinese Population Based on the Development of Mandibular Second and Third Molars.

ABSTRACT: Objective To establish a linear relationship between age and the development process of the mandibular second, third molars in a northern Chinese population and test its accuracy in inferring whether the subject is over 18 years old. Methods The orthopantograms （OPGs） of 960 northern Chinese people aged between 10.00 to 25.99 years were selected. The gender, birth date, and date of the orthopantograms of each sample were recorded. The development stages of left mandibular second and third molars were divided with Demirjian method. The experimental group （760 individuals） constructed a Logistic regression equation to infer whether the subject is over 18 years old. The validation group （200 individuals） tested the accuracy of the equation. Results When only using third molar, the formula obtained for inferring whether the subject's age is over 18 years is： logit P1=-18.276+0.494 x1+2.953 x3. When combining the second and the third molars, the formula is： logit P2=-21.045+3.177 x1+0.619 x2+2.088 x3. （x1 stands for gender, male is marked as 1 and female is marked as 2; x2 and x3 stand for the development stage of second molar and third molar, respectively）. The accuracy rates of this equation in estimating whether ages of subjects from the validation group were over 18 years old for males and females were 88.0%-92.0% and 85.0%-88.0%, respectively. Conclusion The age estimation formulas based on the development of mandibular second and third molars have important value in judging whether the individual is over 18 years old. Meanwhile, the accuracy of age estimation can be improved by combining the development of second and third molars together.

Research Progress of Age Estimation Based on the Demirjian's Method.

ABSTRACT: In recent years, individual age estimation has gained increasing attention in forensic practice. Common age estimation methods mainly use developments of teeth and bones. Compared with bones, teeth are stronger and are highly resistant to external factors, therefore plays an important role in age estimation. Demirjian's method is one of the most commonly used methods to evaluate dental age and has been widely used in many countries. In this paper, the accuracy of its application in different populations is reviewed. It is found that compared with the chronological ages, most of the estimated ages are overestimated. By combining research results of many scholars and by analyzing, it can be assumed that this situation may be related with race, region, sex, etc.

[Research Progress of Age Estimation Based on Age-related Changes of Dentin-pulp Complex].

Age estimation is a hot and difficult issue in forensic practice. Teeth are the most solid organs in human body and can be kept in vitro for a long time. With age, the secondary dentin gradually generates and the volume of pulp cavity constantly decreases. Therefore, forensic dentists proposed that age-related changes of dentin-pulp complex could be used to estimate age, which has been widely applied in forensic practice over the years. Due to the development of imaging technology, a variety of methods have been advocated by forensic dentists to detect the age-related changes of dentin-pulp complex for age estimation. However, different methods have their own advantages and limitations, forensic scientists should combine the use of different methods for improving the accuracy of age estimation according to the actual situation. This paper reviews current research of age estimation based on dentin-pulp complex, so as to provide reference for related research.

Responses of blood glucose, insulin, glucagon, and Fatty acids to intraruminal infusion of propionate in hanwoo.

This study was carried out to investigate the effects of intraruminal infusion of propionate on ruminal fermentation characteristics and blood hormones and metabolites in Hanwoo (Korean cattle) steers. Four Hanwoo steers (average body wt. 270 kg, 13 month of age) equipped with rumen cannula were infused into rumens with 0.0 M (Water, C), 0.5 M (37 g/L, T1), 1.0 M (74 g/L, T2) and 1.5 M (111 g/L, T3) of propionate for 1 hour per day and allotted by 4×4 Latin square design. On the 5th day of infusion, samples of rumen and blood were collected at 0, 60, 120, 180, and 300 min after intraruminal infusion of propionate. The concentrations of serum glucose and plasma glucagon were not affected (p>0.05) by intraruminal infusion of propionate. The serum insulin concentration at 60 min after infusion was significantly (p<0.05) higher in T3 than in C, while the concentration of non-esterified fatty acid (NEFA) at 60 and 180 min after infusion was significantly (p<0.05) lower in the propionate treatments than in C. Hence, intraruminal infusion of propionate stimulates the secretion of insulin, and decreases serum NEFA concentration rather than the change of serum glucose concentration.

Heightened clinical suspicion of pulmonary embolism and disregard of the D-dimer assay: a contemporary trend in an era of increased access to computed tomography pulmonary angiogram?

BACKGROUND: Prospective studies have shown that utilising qualitative D-dimers in those with a low Wells pre-test probability (PTP) of pulmonary embolism (PE) have significantly reduced the number of computed tomography pulmonary angiograms (CTPA) being performed. These studies have been based on a PE prevalence of approximately 6% in the low PTP group. AIM: This study compares the diagnostic approach to PE in the study institution to well-established guidelines. The study also re-examines the cost-benefit analyses of qualitative d-dimers and CTPA in the low PTP group. METHODS: A retrospective study of 169 consecutive CTPA requested in the emergency department of a major teaching hospital during a 12-month period. RESULTS: The prevalence of PE was 0% (0/65), 11.7% (9/77) and 0% (0/2) in the low, moderate and high Wells PTP groups respectively, and 6.3% (9/144) overall. PTP was documented in 10 (6.9%) cases, and the qualitative Clearview Simplify D-dimer was only ordered in 33.8% (22/65) of low PTP subjects. The false positive D-dimer rate was 90.2% (37/41). Cost-benefit analysis and assay performance defines a narrow range of low PTP PE prevalence between 1% and 5% for the utilisation of the qualitative D-dimer assay. CONCLUSIONS: The overall prevalence of PE in subjects undergoing CTPA was significantly lower compared with data in the literature. The authors recommend warranted clinical suspicion of PE should be confirmed by a senior physician prior to placing a patient in the PE work-up pathway. In such patients, the qualitative D-dimer assay should be utilised if PTP is low, and the exclusionary efficiency of the D-dimer will be improved in the setting of higher PE prevalence in this subgroup. Hospitals should audit local PE prevalence, as cost-benefit analyses raises questions about the effectiveness of D-dimers when PE prevalence is very low in the low PTP subgroup.

Efficacy of tomato powder as antioxidant in cooked pork patties.

This study was carried out to determine the effects of tomato powder (TP) on cooked pork patties during storage at 10±1°C in the dark. The total phenolic and flavonoid contents of TP extract were 26.22 mg gallic acid/100 g and 3.52 mg quercetin/100 g, respectively. The extract of TP showed a potential antioxidant activity in the DPPH radical-scavenging assay (EC50 = 16.76 µg/mL). Pork patties were manufactured with 0.25% (T1), 0.5% (T2), 0.75% (T3) and 1.0% (T4) TP in a basic formula (C). The pH and volatile basic nitrogen (VBN) values of T2, T3 and T4 patties were lower (p<0.05) than the C patties during storage. Increased concentration of TP in meat patties decreased (p<0.05) the 2-thiobarbituric acid reactive substances (TBARS) and total plate count (TPC) values at d 7 of storage. Tomato treated-patties had lower (p<0.05) values for lightness (L*), but higher (p<0.05) values for redness (a*) and yellowness (b*) at d 3 and 7 of storage compared with the C. In the case of sensory evaluation, the scores of colour, flavour and overall acceptability of T3 and T4 patties were higher (p<0.05) than those of the C patty after 3 or 7 days of storage.

External validation of AF-BLEED for predicting major bleeding and for tailoring NOAC dose in AF patients: A post hoc analysis in the ENGAGE AF-TIMI 48.

OBJECTIVE: AF-BLEED, a simple bleeding risk classifier, was found to predict major bleeding (MB) in patients with atrial fibrillation (AF) and identify AF patients at high risk of MB who might potentially benefit from a lower direct oral anticoagulant dose. This post hoc study aimed to externally validate these findings in the ENGAGE AF-TIMI 48 (Effective aNticoaGulation with factor Xa next Generation in Atrial Fibrillation-Thrombolysis in Myocardial Infarction study 48) trial. METHODS: The ENGAGE AF-TIMI 48 trial randomized AF patients to higher-dose edoxaban regimen (HDER 60/30 mg) versus lower-dose edoxaban regimen (LDER 30/15 mg), with prespecified dose reduction criteria. AF-BLEED was calculated in the modified intention-to-treat cohort (n = 21,026 patients) used for primary outcome analysis. Annualized event rates and hazard ratios (HRs) were obtained for the primary composite outcome (PCO) and its single components (MB, ischemic stroke/systemic embolism and death) to compare LDER 30 mg with HDER 60 mg in both AF-BLEED classes. RESULTS: AF-BLEED classified 2882 patients (13.7 %) as high-risk, characterized by a two- to three-fold higher MB risk than AF-BLEED classified low-risk patients. AF-BLEED classified high-risk patients randomized to LDER 30 mg demonstrated a 3.3 % reduction in MB at the cost of a 0.5 % increase in ischemic stroke/systemic embolism. LDER 30 mg resulted in a 3.1 % reduction of PCO compared to HDER 60 mg (HR of 0.81; 95%CI 0.65-1.01). Additional to existing dose reduction criteria, another 6 % of patients could potentially benefit of this dose adjustment strategy. CONCLUSION: AF-BLEED could identify AF patients to be at high risk of major bleeding. Our findings support the hypothesis that LDER 30 mg might provide a reasonable option in AF patients with legitimate bleeding concerns.

[Effects of porcine urinary bladder matrix on motility and polarization of bone marrow-derived macrophages in mice].

Objective: To explore the effects of porcine urinary bladder matrix (UBM) on the motility and polarization of bone marrow-derived macrophages in mice, so as to provide evidence for the rational selection of stent in clinical wound repair. Methods: The method of experimental research was used. The microstructure of porcine UBM and absorbable dressing was observed under scanning electron microscope. Polyacrylamide gel electrophoresis was used to observe the protein distribution of the two stent extracts. The primary macrophages were induced from bone marrow-derived cells isolated from six 6-8-week-old male C57BL/6J mice (mouse age, sex, and strain, the same below) and identified. Three batches of macrophages were divided into porcine UBM extract group and absorbable dressing extract group. The cells in each group were cultured with Dulbecco's modified Eagle medium/F12 medium containing the corresponding extracts. The cell migration rate was detected and calculated on 1, 3, and 7 d after scratching by scratch test. The number of migrated cells at 12 and 24 h of culture was detected by Transwell experiment. The percentages of CD206 and CD86 positive cells at 24 h of culture was detected by flow cytometer. The numbers of sample in the above cell experiments were all 3. An incision was prepared on the left and right back of twelve mice, respectively. The left incision of each mouse was included in porcine UBM group and the right incision was included in absorbable dressing group, and the corresponding stents were implanted into the incisions respectively. On post operation day (POD) 7 and 14, the number of inflammatory cells infiltrated in the stent was detected by hematoxylin-eosin staining; the number of F4/80, transforming growth factor-ß1 (TGF-ß1), vascular endothelial growth factor (VEGF), and matrix metalloprotein-9 (MMP-9) positive cells and type Ⅰ collagen deposition in stents were observed by immunohistochemistry; the percentages of F4/80, CD86, and CD206 positive cells were observed by immunofluorescence staining. The numbers of sample in the above animal experiments were all 6. Data were statistically analyzed with analysis of variance for factorial design, analysis of variance for repeated measurement, and independent sample t test. Results: Porcine UBM has a dense basement membrane structure on one side and porous propria containing a fibrous structures on the other. Both sides of the absorbable dressing had three-dimensional porous structure. In the molecular weight range of (50-70)×103, multiple non-type Ⅰ collagen bands appeared in the lanes of porcine UBM extract, while no obvious bands appeared in the lanes of absorbable dressing extract. It had been identified that mouse bone marrow-derived cells had been successfully induced into macrophages. The cell migration rates in porcine UBM extract group were significantly higher than those in absorbable dressing extract group on 1, 3, and 7 d after scratching (with t values of 15.31, 19.76, and 20.58, respectively, P<0.05). The numbers of migrated cells in porcine UBM extract group were significantly more than those in absorbable dressing extract group at 12 and 24 h of culture (with t values of 12.20 and 33.26, respectively, P<0.05). At 24 h of culture, the percentage of CD86 positive cells in porcine UBM extract group ((1.27±0.19)%) was significantly lower than (7.34±0.14)% in absorbable dressing extract group (t=17.03, P<0.05);the percentage of CD206 positive cells in porcine UBM extract group was (73.4±0.7)%, significantly higher than (32.2±0.5)% in absorbable dressing extract group (t=119.10, P<0.05). On POD 7 and 14, the numbers of inflammatory cells infiltrated in the stents in porcine UBM group was significantly more than those in absorbable dressing group (with t values of 6.58 and 10.70, respectively, P<0.05). On POD 7 and 14, the numbers of F4/80, TGF-ß1, VEGF, and MMP-9 positive cells in the stents in porcine UBM group were significantly more than those in absorbable dressing group (with t values of 46.11, 40.69, 13.90, 14.15, 19.79, 32.93, 12.16, and 13.21, respectively, P<0.05); type Ⅰ collagen deposition in the stents in porcine UBM group was more pronounced than that in absorbable dressing group; the percentages of CD206 positive cells in the stents in porcine UBM group were significantly higher than those in absorbable dressing group (with t values of 5.05 and 4.13, respectively, P<0.05), while the percentages of CD86 positive cells were significantly lower than those in absorbable dressing group (with t values of 20.90 and 19.64, respectively, P<0.05), and more M2-type macrophages were seen in the stents in porcine UBM group and more M1-type macrophages were seen in the stents in absorbable dressing group. Conclusions: Porcine UBM can enhance macrophage motility, induce M2 polarization and paracrine function, create a microenvironment containing growth factors such as TGF-ß1 and MMP-9 tissue remodeling molecules, and promote tissue regeneration and extracellular matrix remodeling in mice.

[Management strategy of femoral artery pseudoaneurysm combined with infectious wounds].

Objective: To investigate the surgical treatment methods of femoral artery pseudoaneurysm combined with infectious wounds and to evaluate the clinical effects. Methods: The retrospective observational research method was used. Twelve patients with femoral artery pseudoaneurysm combined with infectious wounds who met the inclusion criteria were admitted to Nanjing University of Chinese Medicine Wuxi Integrated Traditional Chinese and Western Medicine Hospital (Affiliated Hospital of Jiangnan University) from October 2014 to September 2022, including 6 males and 6 females, aged from 46 to 78 years. In the primary operation, debridement, tumor resection, and artery suture/venous grafting to repair the artery/artery ligation were performed, and the wound area after tumor resection ranged from 4.0 cm×1.5 cm to 12.0 cm×6.5 cm. Wounds that could be sutured were treated with tension reduction suture and extracutaneous continuous vacuum sealing drainage (VSD), while large wounds that could not be sutured were treated with VSD to control infection. In the secondary operation, tension reduction suture was performed to repair the wounds that could be sutured; large wounds were repaired with adjacent translocated flaps with area of 9.0 cm×5.0 cm to 15.0 cm×7.0 cm. Additionally, when the length of the exposed femoral artery was equal to or over 3.0 cm, the wounds were repaired with additional rectus femoris muscle flap with length of 15.0 to 18.0 cm. The donor areas of the flaps were directly sutured. The wound with artery ligation was treated with stamp skin grafting and continuous VSD. The bacterial culture results of the wound exudate samples on admission were recorded. The intraoperative blood loss, the location of femoral artery rupture, the artery treatment method, and the wound repair method in the primary operation were recorded, and the durations of catheter lavage, catheter drainage, and VSD treatment, and the drainage volume after the operation were recorded. The repair method of wounds in the secondary operation, the durations of catheter drainage and VSD treatment, and the total drainage volume after the operation were recorded. The survivals of flap/muscle flap/stamp skin grafts were observed, and the wound healing time was recorded. Follow-up after discharge was performed to evaluate the quality of wound healing and the walking function and to check whether the pulsatile mass disappeared. B-ultrasound or computed tomography angiography (CTA) was performed again to observe potential pseudoaneurysm recurrence and evaluate the patency of blood flow of the femoral artery. Results: The bacterial culture results of wound exudate samples of all the patients were positive on admission. The blood loss was 150 to 750 mL in the primary operation. The arterial ruptures were located in the femoral artery in 8 cases, in the external iliac artery in 2 cases, and in the femoral arteriovenous fistula in 2 cases. Six cases received direct artery suture, 4 cases received autologous great saphenous vein grafting to repair the artery, 1 case received autologous great saphenous vein bypass surgery, and 1 case received artery ligation. The primary wound suture was performed in 4 cases, along with catheter lavage for 3 to 5 days, catheter drainage for 4 to 6 days, VSD treatment for 5 to 7 days, and a total drainage volume of 80 to 450 mL after the surgery. In the secondary operation, the wounds were sutured directly in 3 cases along with catheter drainage for 2 to 3 days, the wound was repaired with scalp stamp skin graft and VSD treatment for 5 days in 1 case, the wounds were repaired with adjacent translocated flaps in 2 cases with catheter drainage for 2 to 3 days, and the wounds were repaired with rectus femoris muscle flaps+adjacent translocated flaps in 2 cases with catheter drainage for 3 to 5 days . The total drainage volume after the secondary operation ranged from 150 to 400 mL. All the skin flaps/muscle flaps/skin grafts survived after operation. The wound healing time ranged from 15 to 36 days after the primary operation. Follow-up of 2 to 8 months after discharge showed that the wounds of all patients healed well. One patient who underwent femoral artery ligation had calf amputation due to foot ischemic necrosis, and the rest of the patients regained normal walking ability. The pulsatile mass disappeared in inguinal region of all patients. B-ultrasound or CTA re-examination in 6 patients showed that the blood flow of femoral artery had good patency, and there was no pseudoaneurysm recurrence. Conclusions: Early debridement, tumor resection, and individualized artery treatment should be performed in patients with femoral artery pseudoaneurysm combined with infected wounds. Besides, proper drainage and personalized repair strategy should be conducted according to the wound condition to achieve a good outcome.

Implication of microglia in ketamine-induced long-term cognitive impairment in murine pups.

BACKGROUND: Ketamine, a non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonist, is widely applicable to anesthesia, analgesia, and sedation. However, the function and mechanisms of ketamine in the long-term learning and memory function of neonatal mice are unclear. OBJECTIVE: The present study aims to investigate whether long-term learning and memory function will be affected by multiple ketamine exposures in the early development period. METHODS: The mRNA and protein levels were measured by RT-qPCR and western blot, respectively. The Morris Water Maze test was performed to assess spatial learning and memory. RESULTS: We identified that neonatal exposure to ketamine downsized the positive neurons for microtubule-associated protein doublecortin (DCX) and Ki67 in hippocampal dentate gyrus at the juvenile and late adolescence stages. Double-labeling tests demonstrated that the counts of Iba1+ cells and Ki67+ cells were pronouncedly diminished with exposure to ketamine. Further, qPCR assays to screen the key factors predisposing the populations and maturation of microglia exhibited remarkable decline of CX3CR1 mRNA levels in ketamine group versus the control group. The close relation of microglia to synaptic plasticity was depicted by the significantly downregulated synaptic plasticity-related proteins NR2B and PSD-95 subsequent to multiple exposures to ketamine. Finally, we found that both the protein and mRNA levels of BDNF were markedly decreased in ketamine group versus the control group. CONCLUSION: We found that multiple exposures to ketamine in neonatal mice lead to spatial learning and memory dysfunction. The alterations of microglial development and function are the possible mechanisms of long-term learning and memory impairment.

Determinants of label non-adherence to non-vitamin K oral anticoagulants in patients with newly diagnosed atrial fibrillation.

Aims: To evaluate the extent and determinants of off-label non-vitamin K oral anticoagulant (NOAC) dosing in newly diagnosed Dutch AF patients. Methods and results: In the DUTCH-AF registry, patients with newly diagnosed AF (<6 months) are prospectively enrolled. Label adherence to NOAC dosing was assessed using the European Medicines Agency labelling. Factors associated with off-label dosing were explored by multivariable logistic regression analyses. From July 2018 to November 2020, 4500 patients were registered. The mean age was 69.6 ± 10.5 years, and 41.5% were female. Of the 3252 patients in which NOAC label adherence could be assessed, underdosing and overdosing were observed in 4.2% and 2.4%, respectively. In 2916 (89.7%) patients with a full-dose NOAC recommendation, 4.6% were underdosed, with a similar distribution between NOACs. Independent determinants (with 95% confidence interval) were higher age [odds ratio (OR): 1.01 per year, 1.01-1.02], lower renal function (OR: 0.96 per ml/min/1.73 m2, 0.92-0.98), lower weight (OR: 0.98 per kg, 0.97-1.00), active malignancy (OR: 2.46, 1.19-5.09), anaemia (OR: 1.73, 1.08-2.76), and concomitant use of antiplatelets (OR: 4.93, 2.57-9.46). In the 336 (10.3%) patients with a reduced dose NOAC recommendation, 22.9% were overdosed, most often with rivaroxaban. Independent determinants were lower age (OR: 0.92 per year, 0.88-0.96) and lower renal function (OR: 0.98 per ml/min/1.73 m2, 0.96-1.00). Conclusion: In newly diagnosed Dutch AF patients, off-label dosing of NOACs was seen in only 6.6% of patients, most often underdosing. In this study, determinants of off-label dosing were age, renal function, weight, anaemia, active malignancy, and concomitant use of antiplatelets.

[Influence of porcine urinary bladder matrix and porcine acellular dermal matrix on wound healing of full-thickness skin defect in diabetic mice].

Objective: To compare the difference of pro-healing effect of porcine urinary bladder matrix (UBM) and porcine acellular dermal matrix (ADM) on full-thickness skin defect wounds in diabetic mice. Methods: Thirty-six type 2 diabetic BKS db/db mice aged 10 weeks were divided into UBM group and ADM group according to the random number table, with 18 mice in each group and preoperative molarity of non-fasting blood glucose higher than 16.6 mmol/L. A circular full-thickness skin defect wound with 6 mm in diameter was made on the back of each mouse, and porcine UBM and porcine ADM scaffolds were implanted into the wounds of both groups correspondingly. Immediately after operation and on post operation day (POD) 7, 14, and 28, wounds were observed generally. On POD 7, 14, and 28, 6 mice of each group were collected respectively to calculate the rate of wound epithelialization, and then the corresponding mice were sacrificed after calculation, and the wound tissue was harvested to make slices. Six slices of the mice in the 2 groups on POD 7 and 14 were respectively collected to stain with haematoxylin-eosin (HE), and 6 slices on POD 7 and 28 had Masson's staining, which were used to observe histopathological changes and scaffold degradation. On POD 7 and 14, 24 slices of each mouse in the 2 groups were collected respectively to detect alpha smooth muscle actin (α-SMA) and CD31 positive expression denoting the growth of myofibroblasts and neovessels respectively and observe the distribution and activation of macrophages with immunohistochemical staining. The wound tissue of mice in the 2 groups on POD 7 and 14 was harvested to detect mRNA expressions of fibroblast growth factor 2 (FGF-2), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and transforming growth factor ß(1) (TGF-ß(1)) by real-time fluorescence quantitative reverse transcription polymerase chain reaction. The sample number of above-mentioned indexes in each group at each time point was 6. Data were statistically analyzed with analysis of variance for factorial design, t test, and Bonferroni correction. Results: (1) General observation showed that integration of UBM scaffold into the wounds of mice in UBM group on most time points was superior, and integration of ADM scaffold into the wounds of mice in ADM group on most time points was inferior. On POD 28, epidermis still did not form in some region of scaffold surface of wounds of mice in ADM group, while wounds of mice in UBM group were completely epithelialized. On POD 7, 14, and 28, wound epithelialization rates of mice in UBM group were respectively (22.4±6.4)%, (68.6±12.4)%, and 100.0%, all significantly higher than (4.5±2.2)%, (23.6±4.6)%, and (64.2±13.2)% in ADM group (t=7.427, 9.665, 7.655, P<0.01). (2) HE staining and Masson's staining showed that a large number of cells appeared in wound scaffold of mice in UBM group on POD 7; cells distributed in the whole region of UBM scaffold on POD 14; dermal tissue with structure similar to normal skin formed in the wounds and the fibrous morph of UBM scaffolds disappeared on POD 28. Only a small number of cells appeared in inside of wound scaffolds of mice in ADM group on POD 7; on POD 14, cells were sparsely distributed in ADM scaffolds; on POD 28, the morph of originally robust collagen fiber of ADM scaffolds was still clear and visible. (3) On POD 7, a large number of accumulated myofibroblasts and neovessels appeared in the lower layers of scaffolds of wounds of mice in UBM group; on POD 14, evenly distributed myofibroblasts and neovessels appeared in the upper layers of UBM scaffolds, and most vessels were perfused. On POD 7 and 14, myofibroblasts were sparsely distributed in scaffolds of wounds of mice in ADM group with no or a few neovascular structures perfused unobviously. On POD 7 and 14, α-SMA positive expressions in scaffolds of wounds of mice in UBM group were significantly higher than those in ADM group (t=25.340, 6.651, P<0.01); CD31 positive expressions were also significantly higher than those in ADM group (t=34.225, 10.581, P<0.01). (4) On POD 7, a large number of macrophages appeared in the lower layers of scaffolds of wounds of mice in UBM group; on POD 14, macrophages infiltrated into the internal region of UBM scaffolds, and M2 polarization occured without M1 polarization. On POD 7, a small number of macrophages appeared in the bottom of scaffolds of wounds of mice in ADM group; on POD 14, macrophages were few in internal region of ADM scaffold, and neither M2 polarization nor M1 polarization occurred. (5) On POD 7 and 14, mRNA expressions of FGF-2, VEGF, PDGF, and TGF-ß(1) in the wound tissue of mice in UBM group were all significantly higher than those in ADM group (t=7.007, 14.770, 10.670, 8.939; 7.174, 7.770, 4.374, 4.501, P<0.01). Conclusions: Porcine UBM scaffold is better than porcine ADM in facilitating wound repair and dermis reconstruction of full-thickness skin defects in diabetic mice through the induction of myofibroblasts and macrophages immigration, the promotion of neovascularization and expression of pro-healing growth factors.