Invasive Candida parapsilosis Bloodstream Infections in Children: The Antifungal Susceptibility, Clinical Characteristics and Impacts on Outcomes.

Background: Candida parapsilosis is the most common non-albicans candida species that causes invasive candidiasis, but little is known about its impacts on the outcomes of pediatric patients. We aimed to characterize the clinical characteristics, risk factors and outcomes of C. parapsilosis bloodstream infections (BSIs) in children. Methods: All pediatric patients with Candida parapsilosis BSIs between 2005 and 2020 from a medical center in Taiwan were enrolled and analyzed. The antifungal susceptibility, clinical manifestations, management and outcomes were investigated. Cases of Candida parapsilosis BSIs were compared between patients with C. albicans BSIs and other Candida spp. BSIs. Results: During the study period, 95 episodes (26.0% of total cases) of Candida parapsilosis BSIs were identified and analyzed. No significant difference was found between pediatric patients with C. parapsilosis BSIs and those with C. albicans BSIs in terms of patients' demographics, most chronic comorbidities or risk factors. Pediatric patients with C. parapsilosis BSIs were significantly more likely to have previous azole exposure and be on total parenteral nutrition than those with C. albicans BSIs (17.9 vs. 7.6% and 76.8 vs. 63.7%, p = 0.015 and 0.029, respectively). The duration of C. parapsilosis candidemia was relatively longer, and therefore patients often required a longer duration of antifungal treatment when compared with those of C. albicans candidemia, although the candidemia-attributable mortality rates were comparable. Of the C. parapsilosis isolates, 93.7% were susceptible to all antifungal agents, and delayed appropriate antifungal treatment was an independent factor in treatment failure. Conclusions: Pediatric patients with C. parapsilosis BSIs were more likely to have previous azole exposure and be on total parenteral nutrition, and the clinical significances included a longer duration of candidemia and patients often required a longer duration of antifungal treatment.

Development of a hybrid hydrogel for submucosal injection in endoscopic resection of gastrointestinal neoplasm: From laboratory to clinical trial.

Background and study aims Submucosal injection solution is essential for successful endoscopic resection of the early gastrointestinal tumor. We evaluated a new endoscopic hydrogel for submucosal injection and its clinical feasibility. Patients and methods A hydrogel (AceGel) containing 0.4% sodium alginate and 2% calcium lactate was developed for ex vivo and animal studies. Subsequently, a prospective, single-arm study was conducted to assess its feasibility and safety in humans. Patients with gastrointestinal neoplasms undergoing endoscopic resection were enrolled. All patients underwent endoscopic surveillance after 4 weeks and outpatient follow-up at week 6. Afterward, they received endoscopic follow-up according to the medical routine. Results In the ex vivo experiments, the submucosal elevation height of AceGel was equivalent to sodium hyaluronate and superior to saline or glycerol. Animal studies showed that the excised wounds healed well without surrounding tissue damage. Twelve patients participated in the clinical trial, including three, two, and seven patients with esophageal, gastric, and colonic lesions, respectively. The mean neoplasm size and submucosal injection volumes were 24.0±8.6 mm and 22.8±19.9 mL, respectively. All patients had adequate wound healing on 4-week surveillance endoscopy, and none had serious adverse events during 6-week follow-up. Moreover, endoscopic follow-up showed complete wound healing after 6 to 46 months without local mucosal inflammation in all patients. Conclusions AceGel is good for endoscopic submucosal injection and demonstrated its usefulness in durable mucosal elevation for endoscopic therapy in preclinical tests. This clinical trial shows its safety and feasibility in all participating patients.

Maternal psychological distress in the early postpartum period during COVID-19 pandemic: a pilot study.

BACKGROUND: The coronavirus disease 2019 infection (COVID-19) pandemic is a new global outbreak disease. According to the Taiwan Centers for Diseases Control statement, hospitals had to change their corresponding measures to prevent the spread of COVID-19. The frequency of parental visits to the special care nursery was reduced from three times to once daily. Visiting was not permitted from April 4 to May 10, 2020, and rooming-in with healthy neonates was discontinued, which could increase maternal postpartum distress. Therefore, this study was conducted to determine whether COVID-19 prevention increased maternal psychological distress. METHODS: This prospective study used convenience sampling to enroll healthy mothers who had just delivered via normal spontaneous delivery. Based on the neonates' status and visiting times, mothers were grouped into no-rooming-in, rooming-in, no-visiting, and one-visit/day groups. Mothers' baseline characteristics were compared using the Chi-square or Fisher's exact test and t-test. Salivary cortisol levels and scores of Chinese versions of the Perceived Stress Scale (PSS) and State-Trait Anxiety Inventory were evaluated on postpartum days 1 and 3 and analyzed by one-way analysis of variance and a paired t-test. RESULTS: There were 16, 58, 28, and 47 women categorized as no-rooming-in, rooming-in, no-visit, and one-visit/day groups, respectively. No significant differences were found between groups in mothers' baseline characteristics and postpartum salivary cortisol levels. The PSS on day 3 was significantly higher than on day 1 in every group (p < 0.001). The PSS increasing trend in the no-rooming-in group was significantly greater than that in the no-visit group (p = 0.02) and significantly greater in the rooming-in group than that in the one-visit/day group (p = 0.001). CONCLUSION: Postpartum stress increased for all mothers and was an even more significant response to the COVID-19 pandemic than the stress associated with neonates' hospitalization.

Echocardiographic Determination of Percutaneous Central Venous Catheters in the Superior Vena Cava: A Prospective Cohort Study.

Malposition of percutaneous central venous catheters (PCVCs) in the superior vena cava (SVC) is common. We previously showed that real-time sonography was safer and faster than radiography in identifying PCVC tip location in the inferior vena cava (IVC). However, in preterm infants, determining PCVC tip location in the SVC is complicated by endotracheal or nasogastric tubes in situ and emphysematous lung conditions. We aimed to find an appropriate sonographic view by which to assess PCVC tip location in the SVC compared to the sonographic examination of PCVC in the IVC. Neonates (n = 50) with PCVCs in the SVC were enrolled and their data (gestational age, gender, birth weight, body weight at intervention, repositioning rate, and duration of tip assessment) were compared with retrospective data of 50 neonates with PCVCs in the IVC. The mean gestational age in the groups of IVC and SVC was 31.43 weeks and 32.16 weeks, respectively. The mean birth weight in the groups of IVC and SVC was 1642.18 g and 1792.00 g, respectively. Placement of an S12-4 ultrasound sector transducer to obtain clear parasternal views of the aorta allows visualization of PCVC tips in the SVC and near the cavoatrial junction. PCVC repositioning rates were not significantly different between the two groups (p = 0.092). Sonography examinations in the SVC had a longer duration than those in the IVC (p < 0.001). Sonography provides an accurate method for determining PCVC tip location in the SVC.

Relationship Between Serum Concentration of Adrenomedullin and Myocardial Ischemic T Wave Changes in Patients With Lung Cancer.

Background: Patients with lung cancer are at increased risk for the development of cardiovascular diseases. Molecular markers for early diagnosis of cardiac ischemia are of great significance for the early prevention of cardiovascular events in patients with lung cancer. By evaluating the relationship between adrenomedullin (ADM) and myocardial ischemic T wave changes, the clinical value of circulating ADM as a predictor of myocardial ischemia in patients with lung cancer is confirmed. Methods: We enrolled patients with lung cancer and healthy people from 2019 to 2021 and extracted a detailed ECG parameter. After adjustment for potential confounders, logistic regression was used to assess the association of clinical data. We performed analyses on differences in T wave between patients with lung cancer and healthy people, and the relationship between T wave and ADM among patients with lung cancer. Receiver operator characteristic (ROC) curves were drawn to confirm the diagnostic value of biomarkers. Results: After adjusting for potential confounders, the incidence of T wave inversion or flattening in patients with lung cancer was higher than in healthy people (OR: 3.3228, P = 0.02). Also, further analysis of the data of lung cancer patients revealed that the ADM in lung cancer patients with T wave inversion or flat was higher than those with normal T wave (189.8 ± 51.9 vs. 131.9 ± 38.4, p < 0.001). The area under the ROC curve was 0.8137. Conclusion: Among the patients with lung cancer, serum ADM concentration is associated with the incidence of the abnormal T wave. ADM might be a potentially valuable predictor for heart ischemia in patients with lung cancer.

Comparison of Different Endoscopic Methods Used for Managing Choledocholithiasis in Patients with End-Stage Renal Disease Undergoing Hemodialysis.

BACKGROUND AND AIM: Endoscopic sphincterotomy (EST), endoscopic papillary balloon dilation (EPBD), and endoscopic sphincterotomy plus balloon dilation (ESBD) are all techniques used to manage choledocholithiasis. We aim to analyze the efficacy and safety of these techniques for treating choledocholithiasis in patients undergoing hemodialysis (HD). METHODS: We performed a retrospective study of 80 patients with end-stage renal disease (ESRD) on HD who underwent endoscopic retrograde cholangiopancreatography (ERCP) for choledocholithiasis management between August 1st, 2012, and December 31st, 2020, at a medical center in southern Taiwan. These patients were divided into three groups: EST (n = 21), EPBD (n = 28), and ESBD (n = 31). Post-ERCP complications, including pancreatitis, bleeding, cholangitis, and perforation, were reviewed for analysis. RESULTS: There were no significant among-group differences in the rate of complete stone clearance and hospitalization day after ERCP. Patients in the EST group had a higher post-ERCP complication rate than was the case in the other groups (p = 0.016). ESBD significantly reduced post-ERCP bleeding, compared with that occurring with EST (OR 0.07; 95% CI, 0.01-0.72, p = 0.026). There were no significant among-group differences in the rates of pancreatitis and cholangitis. There were no ERCP-related perforations or deaths in this study. CONCLUSIONS: EST, EPBD, and ESBD are efficient methods for treating choledocholithiasis in ESRD patients. ESBD was found to lead to a lower risk of bleeding than EST, and the rate of pancreatitis or cholangitis was comparable for EST and EPBD. Our results suggest that ESBD is the best choice of treatment of choledocholithiasis in patients with ESRD undergoing HD.

Alternate Partial Root-Zone Drip Nitrogen Fertigation Reduces Residual Nitrate Loss While Improving the Water Use but Not Nitrogen Use Efficiency.

The efficient utilization of irrigation water and nitrogen is of great importance for sustainable agricultural production. Alternate partial root-zone drip irrigation (APRD) is an innovative water-saving drip irrigation technology. However, the coupling effects of water and nitrogen (N) supply under APRD on crop growth, water and N use efficiency, as well as the utilization and fate of residual nitrates accumulated in the soil profile are not clear. A simulated soil column experiment where 30-40 cm soil layer was 15NO3-labeled as residual nitrate was conducted to investigate the coupling effects of different water [sufficient irrigation (W1), two-thirds of the W1(W2)] and N [high level (N1), 50% of N1 (N2)] supplies under different irrigation modes [conventional irrigation (C), APRD (A)] on tomato growth, irrigation water (IWUE) and N use efficiencies (NUE), and the fate of residual N. The results showed that, compared with CW1N1, AW1N1 promoted root growth and nitrogen absorption, and increased tomato yield, while the N absorption and yield did not vary significantly in AW2N1. The N absorption in AW2N2 decreased by 16.1%, while the tomato yield decreased by only 8.8% compared with CW1N1. The highest IWUE appeared in AW2N1, whereas the highest NUE was observed in AW2N2, with no significant difference in NUE between AW2N1 and CW1N1 at the same N supply level. The 15N accumulation peak layer was almost the same as the originally labeled layer under APRD, whereas it moved 10-20 cm downwards under CW1N1. The amount of 15N accumulated in the 0-40 cm layer increased with the decreasing irrigation water and nitrogen supply, with an increase of 82.9-141.1% in APRD compared with that in CW1N1. The utilization of the 15N labeled soil profile by the tomato plants increased by 9-20.5%, whereas the loss rate of 15N from the plant-soil column system decreased by 21.3-50.1% in APRD compared with the CW1N1 treatment. Thus, APRD has great potential in saving irrigation water, facilitating water use while reducing the loss of residual nitrate accumulated in the soil profile, but has no significant effect on the NUE absorbed.

Association of furosemide or hydrochlorothiazide use with risk of atrial fibrillation post pacemaker implantation among elderly patients.

BACKGROUND: Atrial fibrillation (AF) induced by artificial pacing is directly related to atrial remodeling. Previous basic research has shown that furosemide aggravates pathologic myocardial remodeling while hydrochlorothiazide alleviates it. However, whether furosemide or hydrochlorothiazide plays a role in developing AF after pacemaker implantation remains unknown. The study aims to investigate the association between oral furosemide or hydrochlorothiazide and the risk of developing AF after pacemaker implantation. METHODS: After a review of electronic medical records, elderly patients with pacemaker implantation and without a known baseline history of AF were included and information on their use of daily oral furosemide or hydrochlorothiazide was extracted. New incident AF cases were confirmed via the records of outpatient visits. A Cox proportional-hazards model was used to evaluate the association between daily oral furosemide or hydrochlorothiazide and risk of developing AF after pacemaker implantation, after adjustment for potential confounders. RESULTS: Among a total of 551 patients aged more than 65 years, 157 AF cases were identified after pacemaker implantation during a maximum follow up of 3.0±1.6 years. Of these, 242 had used furosemide and 97 had used hydrochlorothiazide therapy. Patients taking daily oral furosemide had a relatively higher risk of AF after pacemaker implantation [hazard ratio (HR): 1.507, 95% confidence interval (CI): 1.036-2.192; P=0.032] after being adjusted for related disease and prescribed medications, while oral taking of hydrochlorothiazide was shown to be a non-effective factor (HR: 0.666, 95% CI: 0.413-1.074), which had no statistical significance. CONCLUSIONS: Daily oral furosemide might increase the risk of developing AF after pacemaker implantation in elderly patients, while hydrochlorothiazide has no detrimental effect.

Case Report: Successful Use of Biliary Stent for Iatrogenic Esophageal Perforation Following Balloon Dilation in a 7-Month-Old Infant.

Esophageal perforation is a rare but critical emergency that requires early detection and prompt management. In the pediatric population, iatrogenic injury is the most common etiology of esophageal perforation, and the majority of cases come from stricture dilation. Treatment options include medical management, endoscopic therapy, and surgery. Usually, conservative treatment is appropriate in most carefully selected patients, especially in the setting of early diagnosis and with the absence of severe sepsis. A surgical approach is reserved for a large tear with mediastinum contamination, or clinical deterioration after unsuccessful conservative management. With the advancement of the endoscopy technique, endoscopy therapy using esophageal stents is an available choice for adult populations who have a complicated protracted healing course or comorbidities precluding surgical attempts. However, this procedure is seldom implemented in children, especially in young infants, owing to unavailable equipment and experts. We report our successful use of a fully-covered self-expandable metal biliary stent in managing esophageal perforation in a seven-month-old infant. In light of this encouraging achievement, this model can be applied to more children who have the same problem.

Nonalcoholic fatty liver disease impacts the control of the international normalized ratio in patients with atrial fibrillation.

BACKGROUND: It is unclear whether the therapeutic effect of warfarin in patients with atrial fibrillation (AF) and normal liver function differs between those with and without nonalcoholic fatty liver disease (NAFLD). With this in mind, we aimed to evaluate the impact of NAFLD on the international normalized ratio (INR) control in warfarin-treated AF patients with normal liver function. METHODS: We enrolled 600 AF patients aged 28-94 (median 68) with normal liver function who were receiving daily warfarin therapy, 172 with NAFLD and 428 without. The INR and INR/warfarin dosage rate were measured. Four nested multivariable linear regression models adjusted for potential confounders were used to assess whether there were differences in INR and INR/warfarin dose rate between patients with and without NAFLD. RESULTS: The INR, the percentage of patients with INR within the target range of 2.0-3.0, and the INR/warfarin dose rate were lower in patients with NAFLD than those without. In the maximally adjusted multivariable linear regression models, the INR in NAFLD patients (0.22±0.07, P=0.003) was lower than in non-NAFLD patients, and the INR/warfarin dose rate was slightly lower (0.09±0.06, P=0.10) in NAFLD than in non-NAFLD patients. CONCLUSIONS: Our findings suggest that among AF patients, the therapeutic effect of warfarin is impaired in patients who have NAFLD. Therefore, a slightly higher or personally optimized dosage of warfarin might be necessary among AF patients with NAFLD in order to achieve the INR target range.

Computed tomography classification of endoscopic retrograde cholangiopancreatography-related perforation.

Endoscopic retrograde cholangiopancreatography (ERCP)-related perforation leads to high morbidity and mortality. The Stapfer classification divides patients with different perforation locations and suggests management accordingly. The classification may be unknown if perforation is not detected during endoscopy. We classified patients with ERCP-related perforation (ERP) through computed tomography (CT) and observed the clinical outcomes with varyingly invasive management. Fifty-two cases of ERP between July 2009 and December 2017 were retrospectively reviewed. Of them, 41 who underwent CT for ERCP were included. According to their CT findings, we divided patients into air-alone (n = 16), air-fluid (n = 18), and fluid-alone (n = 7) groups. Perforation severity was graded using the Clavien-Dindo classification for surgical complications. Demographic data and clinical outcomes among different groups were analyzed. Fifteen patients (37%) had an unknown Stapfer classification. More than half of the patients in the air-fluid group had a Clavien-Dindo complication grade of >3. Four patients underwent surgical repair; all of them were from the air-fluid group. All patients in the air- and fluid-alone groups underwent medical treatment without need for subsequent salvage surgery. The air-fluid group had the longest mean hospital stay (25.1 ± 21.9 days) and the exclusive two mortality cases in this study. Patients with ERCP can be divided into groups with different outcomes according to the presence of air or fluid on CT images. Because patients with both air and fluid have the worst clinical outcome, they may require more aggressive treatment than patients with either air or fluid alone.

Elevated level of miR-17 along with decreased levels of TIMP-1 and IL-6 in plasma associated with the risk of in-stent restenosis.

In-stent restenosis is highly related to the deposition of inflammatory extracellular matrix and the migration of endothelial and vascular smooth muscle cells. The miR-17/TIMP-1/interleukin pathway regulates vascular matrix remodeling and plays an important role in the inflammatory reaction. This study identified miR-17 and its related biomarkers in serum that potentially indicated susceptibility to in-stent restenosis (ISR) after coronary artery stenting. Subjects were 42 patients with single de novo coronary artery lesions who underwent regular coronary angiography one year after percutaneous coronary intervention. The clinical baseline information was recorded. Serum levels of biomarkers (including miR-17, TIMP-1, IL-6, IL-8, IL-2R, TNF-alpha, IL-10, and IL-1beta) were measured with real-time PCR or ELISA. Intergroup comparisons were used to compare patients with or without ISR. Compared to levels in the non-restenosis group, the serum miR-17 level was significantly higher (3.13 ± 0.22 vs. 1.06 ± 0.04, p < 0.01) and the serum TIMP-1 and IL-6 levels were significantly lower in the ISR group (TIMP-1: 0.33 ± 0.04 vs. 1.00 ± 0.05, p < 0.01; IL-6: 1.64 ± 0.18 vs. 3.52 ± 0.11, p < 0.01). Moreover, the levels of TIMP-1 and IL-6 decreased as the level of miR-17 increased. Spearman's correlation analysis indicated that the miR-17 level was inversely correlated with TIMP-1 and IL-6 levels. Findings suggest that an elevated level of miR-17 and decreased levels of TIMP-1 and IL-6 may be associated with the risk of ISR, which is in accordance with vascular matrix remodeling and an inflammatory reaction during the pathologic process of ISR. This study highlighted the potential for miR-17, TIMP-1, and IL-6 to serve as biomarkers for ISR.

Rational redesign of a cation···π···π stacking at cardiovascular Fbw7-Skp1 complex interface and its application for deriving self-inhibitory peptides to disrupt the complex interaction.

The Fbw7-Skp1 complex is an essential component in the formation and development of the mammalian cardiovascular system; the complex interaction is mediated through binding of Skp1 C-terminal peptide (qGlu-peptide) to the F-box domain of Fbw7. By visually examining the crystal structure, we identified a typical cation ···π···π stacking system at the complex interface, which is formed by the Trp1159 residue of qGlu-peptide with the Lys2299 and His2359 residues of Fbw7 F-box domain. Both hybrid quantum mechanics/molecular mechanics (QM/MM) analysis of the real domain-peptide complex and electron-correlation ab initio calculation of the stacking system model suggested that the cation···π···π plays an important role in stabilizing the complex; substitution of peptide Trp1159 residue with aromatic Phe and Tyr would not cause a considerable effect on the configuration and energetics of cation···π···π stacking system, whereas His substitution seems to largely destabilize the system. Subsequently, the qGlu-peptide was stripped from the full-length Skp1 protein to define a so-called self-inhibitory peptide, which may rebind to the domain-peptide complex interface and thus disrupt the complex interaction. Fluorescence polarization (FP) assays revealed that the Trp1159Phe and Trp1159Tyr variants have a comparable or higher affinity (K d = 41 and 62 µM) than the wild-type qGlu-peptide (K d = 56 µM), while the Trp1159His mutation would largely impair the binding potency of qGlu-peptide to Fbw7 F-box domain (K d = 280 µM), confirming that the cation···π···π confers both affinity and specificity to the domain-peptide recognition, which can be reshaped by rational molecular design of the nonbonded interaction system. Graphical abstract Stereoview of the complex structure of Fbw7 with Skp1 (PDB: 2ovp), where the Trp1159 residue of Skp1 qGlu-peptide can form a cation···π···π stacking system with the Lys2299 and His2359 residues of Fbw7 F-box domain.

Proposal of a diagnostic algorithm for intraductal ultrasonography to distinguish between benign and malignant biliary strictures.

To reexamine the recognizability of intraductal ultrasonography (IDUS) findings from an imaging database and propose a novel algorithm for clinical application. IDUS images of 102 patients who had undergone IDUS examinations for indeterminate causes of common bile duct dilation were independently reviewed by two endoscopists. The strength of the inter-rater agreement between the endoscopists was analyzed using Cohen's kappa (κ). An algorithm was implemented by arranging the IDUS characteristics according to their recognizability. The proposed algorithm was evaluated by examining the inter-rater agreement and diagnostic accuracy before and after the use of the algorithm. The strength of the inter-rater agreement was good for common bile duct stones with or without acoustic shadowing; intraluminal tumors; or bile duct wall thicknesses of more than or equal to 9 mm (κ > 0.8); followed by intraluminal hypoechoic nodules without common bile duct stone characteristics (κ = 0.771); and finally eccentric wall thickening, outer layer disruption, irregular mucosa, and destructed mural layers (κ: 0.595-0.419). Our algorithm improved the strength of inter-rater agreement with a diagnostic accuracy of 81.4%. We proposed an algorithm according to the recognizability of IDUS characteristics, and it can be used by endoscopists to evaluate such characteristics and determine the cause of biliary obstruction.

Prolonged P-wave duration is associated with atrial fibrillation recurrence after radiofrequency catheter ablation: A systematic review and meta-analysis.

BACKGROUND/OBJECTIVES: A change in the P-wave duration (PWD) was made in patients with atrial fibrillation recurrence (AFR) after radiofrequency catheter ablation (RFCA). The correlation between the PWD and AFR and the potential utilization of a prolonged PWD in the prediction of AFR after RFCA remains unclear. METHODS: Studies of PWD in patients with or without AFR after RFCA, and studies concerned with the predictive effect of prolonged PWD on AFR after RFCA, were included. Outcome measures are reported as absolute risk differences with 95% confidence intervals. A receiver operating characteristic (ROC) curve was used to evaluate the potential cutoff value of PWD for AFR. Summary receiver operating characteristic curve (SROC) analysis was performed to show the overall predictive value of PWD for AFR. RESULTS: Nine studies were included. The overall effect test based on 8 studies that contained a total cohort of 1010 patients showed a highly significant association between prolonged PWD and AFR after RFCA (Z=14.20, P<0.000). A summary that included 4 studies with a combined total of 593 patients indicated a higher risk of AFR in patients with prolonged PWD (Z=5.86, P<0.000). ROC curve analysis indicated 149.5ms as the potential cutoff value of PWD for AFR after RFCA. SROC analysis suggested an acceptable predictive efficiency of PWD for AFR (AUC=0.66). CONCLUSIONS: The risk of AFR after RFCA is strongly related to a prolonged PWD. PWD is one potential low-cost and feasible predictor of AFR after RFCA.

[Treatment of nonunion of femoral shaft fracture after initial locked intramedullary fixation with plate and xenogenic bony plate].

OBJECTIVE: To investigate the clinical effects of plate and xenogenic bony plate for the treatment of nonunion of femoral shaft fracture after initial fixation with locked intramedullary nailing. METHODS: From February 2006 to June 2013, 21 cases with nonunion of femoral shaft fracture after initial fixation with locked intramedullary nailing were treated with femoral plate and contralateral xenogenic bony plate. There were 12 males and 9 females with an average age of 34.8 years old (ranging from 18 to 62 years). The time of nonunion was 9 to 18 months (averaged 12.8 months). The clinical outcomes of the treatment were evaluated by Merchan assessment. RESULTS: All of the patients were primary healing, and on complications such as infection,fat embolism, internal fixation breaking or rotational deformity, shortening were occurred. All the cases were followed up for 13.2 months (ranging from 8 to 24 months). Nineteen cases were bone healed,the time of union averaged 6.2 months (ranging from 4 to 9 months). Two cases appeared delayed union and gained bony union after 7 to 13 months' observation. According to the criterion of Merchan,the results were excellent in 13 cases, good in 6 cases, poor in 2 cases. CONCLUSION: Treatment of nonunion of femoral shaft fracture after initial locked intramedullary fixation with plate and xenogenic bony plate has advantages of high curative rate and low complications, good postoperative function recovery, it is a reliable treatment to elevate the stability of fixation and promote the osteogenesis.

MicroRNA-223 displays a protective role against cardiomyocyte hypertrophy by targeting cardiac troponin I-interacting kinase.

BACKGROUND/AIMS: MicroRNAs play regulatory role in cardiovascular disease. MicroRNA-223 (miR-223) was found to be expressed abundantly in myocardium. TNNI3K, a novel cardiac troponin I (cTnI)-interacting and cardiac hypertrophy related kinase, is computationally predicted as a potential target of miR-223. This study was designed to investigate the cellular and molecular effects of miR-223 on cardiomyoctye hypertrophy, focusing on the role of TNNI3K. METHODS: Neonatal rat cardiomyocytes (CMs) were cultured, and CMs hypertrophy was induced by endothelin-1 (ET-1). In vivo cardiac hypertrophy was induced by transverse aorta constriction (TAC) in rats. Expression of miR-223 in CMs and myocardium was detected by real-time PCR (RT-PCR). MiR-223 and TNNI3K were overexpressed in CMs via chemically modifed sense RNA (miR-223 mimic) transfection or recombinant adenovirus infection, respectively. Cell size was measured by surface area calculation using fluorescence microscopy after anti-α-actinin staining. Expression of hypertrophy-related genes was detected by RT-PCR. The protein expression of TNNI3K and cTnI was determined by Western blots. Luciferase assay was employed to confirm the direct binding of miR-223 to the 3'UTR of TNNI3K mRNA. Intracellular calcium was measured by sensitive fluorescent indicator (Furo-2). Video-based edge detection system was employed to measure cardiomyocyte contractility. RESULTS: MiR-223 was downregulated in ET-1 induced hypertrophic CMs and in hypertrophic myocardium compared with respective controls. MiR-223 overexpression in CMs alleviated ET-1 induced hypertrophy, evidenced by smaller cell surface area and downregulated ANP, α-actinin, Myh6 and Myh7 expression. Luciferase reporter gene assay showed that TNNI3K serves as a direct target gene of miR-223. In miR-223-overexpressed CMs, the protein expression of TNNI3K was significantly downregulated. MiR-223 overexpression also rescued the upregulated TNNI3K expression in hypertrophic CMs. Furthermore, cTnI phosphorylation was downregulated post miR-223 overexpression. Ad.rTNNI3K increased intracellular Ca(2+) concentrations and cell shortening in CMs, while miR-223 overexpression significantly rescued these hypertrophic effects. CONCLUSION: By direct targeting TNNI3K, miR-223 could suppress CMs hypertrophy via downregulating cTnI phosphorylation, reducing intracellular Ca(2+) and contractility of CMs. miR-223 / TNNI3K axis may thus be major players of CMs hypertrophy.

Role of miR-145 in cardiac myofibroblast differentiation.

Following a myocardial infarction (MI), fibroblasts differentiate to myofibroblasts, which possess some of the characteristics of smooth muscle cells (SMCs) and contribute to wound healing. Previous studies suggested that the miR-143/-145 cluster plays a critical role in SMC differentiation. Therefore, we determined whether miR-145 promoted differentiation of cardiac fibroblasts to myofibroblasts. Following coronary occlusion in mice, myocardial miR-145 expression was downregulated at 3 days but was restored at 7 days. In vitro studies showed that hypoxia also downregulated miR-145 in cardiac fibroblasts. The number of α-smooth muscle actin (α-SMA) positive cells in fibroblast cultures was employed to determine their transdifferentiation to cardiac myofibroblasts and was increased by 73.5% after transient transfection with miR-145. Ultrastructural analysis of α-SMA stress fibers revealed that ~95% of the α-SMA(+) cells treated with miR-145 organized their actin-filament bundles with a specific orientation compared to only 15% in the scrambled control group. This orientation of the SMA bundles and their integration with the filamentous actin fibers of the cytoskeleton permit infarct wound contraction. Structural and functional studies showed that miR-145 induced a myofibroblast phenotype, and miR-145 also potentiated the production of mature collagen by myofibroblasts. Repression of KLF5, a target of miR-145, was validated by a chimeric luciferase construct tagged with the full-length 3'-UTR of KLF5. A dramatic decrease in KLF5 and a corresponding increase in myocardin expression were observed after transfecting cultured fibroblasts with miR-145. Similar results were found in vivo: the transient decrease in miR-145 expression 3 days post-MI was associated with an increase in KLF5 and a decrease in myocardin. In addition, in vivo delivery of a miR-145 antagomir 1 day prior to and 2 and 6 days after MI decreased myofibroblast formation and increased scar size. The antagomir also reversed the suppressed expression of KLF5 protein in the scar region at day 7 after MI. In summary, we describe a novel association between miR-145 and fibroblast differentiation toward myofibroblasts. These observations provide a new approach to promote endogenous scar healing and contracture by stimulating the transdifferentiation of cardiac fibroblasts to myofibroblasts.

ERK5 knock down aggravates detrimental effects of hypothermal stimulation on cardiomyocytes via Bim upregulation.

Mechanism of cold induced myocardial injury remained unclear. Our study investigated the role of ERK5/Bim pathway in hypothermal stimulation-induced apoptosis or damage of cardiomyocytes (CMs). Results showed that in CMs which under hypothermal stimulation, ERK5 siRNA promoted expression of Bim protein. Bim siRNA did not influence ERK5 expression but attenuated production of p-ERK5. ERK5 siRNA induced higher apoptosis rate; intracellular Ca(2+) overload; ROS activity; ΔΨm damage in hypothermia stimulated CMs, when compared with hypothermal stimulation solely treated group, while Bim siRNA effected oppositely and canceled pro-apoptotic effect of ERK5 siRNA. In conclusion, ERK5 knock down releases inhibition to Bim expression, induces aggravated apoptosis in CMs under hypothermal stimulation, which related to higher intracellular Ca(2+) overload, ROS activity, and more severe ΔΨm damage. Results revealed regulative role of ERK5/Bim pathway in hypothermal stimulation-induced injure or apoptosis of cardiomyocytes.

The copolymer of Poly(2-dimethylaminoethyl methacrylate) and methacrylated chondroitin sulfate with low cytotoxicity for gene delivery.

Poly(2-dimethylaminoethyl methacrylate) (PDMAEMA) is one of the most potent synthetic nonviral gene-delivery vectors because of its high transfection efficiency. However, the cytotoxicity of PDMAEMA is a major concern for its clinical applications. An anionic crosslinker is synthesized based on a natural polysaccharide, chondroitin sulfate (CS), by introducing methacrylate groups (CSMA). By systematically adjusting the substitution degree of methacrylation on CS and the weight percent of CSMA and PDMAEMA, sol-type copolymers are obtained as a gene-delivery vector. The combination of CS and PDMAEMA is expected not only to reduce the cytotoxicity of PDMAEMA, but also to facilitate better transfection efficiency than PDMAEMA because of the recognition of CS by CD44 receptors on cell surfaces. Two CSMA-modified PDMAEMA copolymers with different CSMA constituents are selected and their polyplexes prepared with plasmid DNA. The cytotoxicity and gene transfection efficiency of the polyplexes are tested and compared with those of PDMAEMA/pDNA. The copolymers of CSMA and PDMAEMA show significantly improved cell viability as compared with PDMAEMA. Their formed polyplexes with pDNA also show lower cytotoxicity than does PDMAEMA/pDNA. The transfection efficiency remarkably increases as the CSMA-modified PDMAEMA/pDNA polyplex is prepared at a weight ratio of 2.4. The internalization mechanism of CSMA-modified PDMAEMA/pDNA in HEK 293T cells is mainly based on caveolae-mediated endocytosis. However, both caveolae-mediated and CD44-mediated endocytosis mechanisms are involved in U87 cells.