Different Cell Responses to Hinokitiol Treatment Result in Senescence or Apoptosis in Human Osteosarcoma Cell Lines.

Hinokitiol is a tropolone-related compound isolated from the heartwood of cupressaceous plants. It is known to exhibit various biological functions including antibacterial, antifungal, and antioxidant activities. In the study, we investigated the antitumor activities of hinokitiol against human osteosarcoma cells. The results revealed that hinokitiol treatment inhibited cell viability of human osteosarcoma U-2 OS and MG-63 cells in the MTT assay. Further study revealed that hinokitiol exposure caused cell cycle arrest at the S phase and a DNA damage response with the induction of γ-H2AX foci in both osteosarcoma cell lines. In U-2 OS cells with wild-type tumor suppressor p53, we found that hinokitiol exposure induced p53 expression and cellular senescence, and knockdown of p53 suppressed the senescence. However, in MG-63 cells with mutated p53, a high percentage of cells underwent apoptosis with cleaved-PARP expression and Annexin V staining after hinokitiol treatment. In addition, up-regulated autophagy was observed both in hinokitiol-exposed U-2 OS and MG-63 cells. As the autophagy was suppressed through the autophagy inhibitor chloroquine, hinokitiol-induced senescence in U-2 OS cells was significantly enhanced accompanying more abundant p53 expression. In MG-63 cells, co-treatment of chloroquine increased hinokitiol-induced apoptosis and decreased cell viability of the treated cells. Our data revealed that hinokitiol treatment could result in different cell responses, senescence or apoptosis in osteosarcoma cell lines, and suppression of autophagy could promote these effects. We hypothesize that the analysis of p53 status and co-administration of autophagy inhibitors might provide more precise and efficacious therapies in hinokitiol-related trials for treating osteosarcoma.

Bilateral pulmonary embolism without deep venous thrombosis was observed after knee arthroscopy: a case report.

BACKGROUND: Symptomatic pulmonary embolism (PE) after knee arthroscopy is extremely rare. If the embolism is not treated promptly, the patient may die. Bilateral pulmonary embolism with associated pulmonary infarct without concomitant deep vein thrombosis has never been reported following routine knee arthroscopy. CASE PRESENTATION: A 50-year-old female patient with no other risk factors other than hypertension, obesity, varicose veins in the ipsilateral lower extremities and elevated triglyceride (TG) presented to our ward. She had experienced sudden chest tightness, polypnea and fainting after going to the bathroom the morning of the second postoperative day and received emergency medical attention. Colour ultrasonography of the extremities showed no deep vein thrombosis. Lung computed tomography angiography (CTA) showed multiple embolisms scattered in both pulmonary artery branches. Thus, emergency interventional thrombolysis therapy was performed, followed by postoperative symptomatic treatment with drugs with thrombolytic, anticoagulant and protective activities. One week later, lung CTA showed a significant improvement in the PEs compared with those in the previous examination. Since the aetiology of PE and no obvious symptoms were discerned, the patient was discharged. CONCLUSION: Although knee arthroscopy is a minimally invasive and quick procedure, the risk factors for PE in the perioperative period should be considered and fully evaluated to enhance PE detection. Moreover, a timely diagnosis and effective treatment are important measures to prevent and cure PE after knee arthroscopy. Finally, clear guidelines regarding VTE thromboprophylaxis following knee arthroscopy in patients with a low risk of VTE development are needed.

Extracellular heat shock protein HSC70 protects against lipopolysaccharide-induced hypertrophic responses in rat cardiomyocytes.

We have recently shown that exogenous administration of extracellular heat shock protein HSC70, a previously recognized intracellular chaperone protein, can protect against LPS-induced cardiac dysfunction through anti-inflammatory actions. However, whether it can also exert anti-hypertrophic effect is unknown. The present study was aimed to investigate the efficacy of HSC70 against cardiac hypertrophy and its underlying molecular mechanisms. Cardiomyocytes were isolated from the cardiac ventricles of neonatal Wistar rats and LPS (1 µg/mL) was used to induce the hypertrophic responses. We found that HSC70 (0.1, 1 and 5 µg/mL) pretreatment attenuated LPS-induced cardiomyocyte hypertrophy dose-dependently. In addition, HSC70 mitigated LPS-induced inflammatory mediators including TNF-α, IL-6, NO, iNOS and COX-2, with down-regulated protein expression of MMP-2 and MMP-9. Moreover, HSC70 repressed LPS-induced signaling of MAPK and Akt. Finally, HSC70 inhibited NF-κB subunit p65, and the DNA binding activity of NF-κB. Taken together, these findings suggest that in vitro HSC70 can exert anti-hypertrophic effects through inhibition of pro-inflammatory mediators, which are potential mediated by the down-regulation of MAPK, Akt and NF-κB signaling pathways. In conclusion, extracellular HSC70 may be a novel pharmacologic strategy in the management of cardiac hypertrophy.

The First Conducting Spin-Crossover Compound Combining a MnIII Cation Complex with Electroactive TCNQ Demonstrating an Abrupt Spin Transition with a Hysteresis of 50†K.

We present herein the synthesis, crystal structure, and electric and magnetic properties of the spin-crossover salt [Mn(5-Cl-sal-N-1,5,8,12)]TCNQ1.5 ⋅2 CH3 CN (I), where 5-Cl-sal-N-1,5,8,12=N,N'-bis(3-(2-oxy-5-chlorobenzylideneamino)propyl)-ethylenediamine, containing distinct conductive and magnetic blocks along with acetonitrile solvent molecules. The MnIII complex with a Schiff-base ligand, [Mn(5-Cl-sal-N-1,5,8,12)]+ , acts as the magnetic unit, and the π-electron acceptor 7,7,8,8-tetracyanoquinodimethane (TCNQ- ) is the conducting unit. The title compound (I) exhibits semiconducting behavior with room temperature conductivity σRT ≈1×10-4  ohm-1 cm-1 and activation energy Δ ≈0.20 eV. In the temperature range 73-123 K, it experiences a hysteretic phase transition accompanied by a crossover between the low-spin S=1 and high-spin S=2 states of MnIII and changes in bond lengths within the MnN4 O2 octahedra. The pronounced shrinkage of the basal Mn-N bonds in I at the spin crossover suggests that the d x 2 - y 2 orbital is occupied/deoccupied in this transition. Interestingly, the bromo isomorphic counterpart [Mn(5-Br-sal-N-1,5,8,12)]TCNQ1.5 ⋅2 CH3 CN (II) of the title compound evidences no spin-crossover phenomena and remains in the high-spin state in the temperature range 2-300 K. Comparison of the chloro and bromo compounds allows the thermal and spin-crossover contributions to the overall variation in bond lengths to be distinguished. The difference in magnetic behavior of these two salts has been ascribed to intermolecular supramolecular effects on the spin transition. Discrete hydrogen bonding exists between cations and cations and anions in both compounds. However, the hydrogen bonding in the crystals of II is much stronger than in I. The relatively close packing arrangement of the [Mn(5-Br-sal-N-1,5,8,12)]+ cations probably precludes their spin transformation.

Predicting severe enterovirus 71 infection: Age, comorbidity, and parental behavior matter.

OBJECTIVE: Enterovirus 71 (EV71) is one of the major pathogens that cause severe enteroviral infections. Our aim was to study the behavioral and household risk factors for its serious complications. METHODS: Between May 2011 and November 2012, we enrolled children who had symptoms of EV71 infection from six hospitals in Taiwan. The caregivers of each patient were interviewed to determine their hand hygiene habits in relation to EV71 infection. The severity of EV71 infection was classified as follows: Stage 1, hand-foot-mouth disease or herpangina; Stage 2, meningitis or myoclonic jerk; Stage 3A, encephalitis; Stage 3B, cardiopulmonary failure. Stages 2 to 3B were defined as severe EV71 infection. Children with Stages 3A and 3B infection were designated as the critical group. RESULTS: A total of 399 patients had laboratory-confirmed EV71 infection. Three risks factors were associated with the different degrees of severity in EV71 infection. Children <2 years old had much greater risks for severe EV71 infection [odds ratio (OR) 1.8; 95% confidence interval (CI), 1.2-2.8], delayed medical evaluation for critical infection (OR 9.4; 95% CI, 3.6-24.1), and developmental retardation for cardiopulmonary failure (OR 8.3; 95% CI, 2.0-33.7). Among all the habits and household factors, caregivers in the critical group had a significantly lower rate in terms of cleaning the faucet after washing their hands (OR 2.63; 95% CI, 1.14-6.08). CONCLUSIONS: Children <2 years old, developmental retardation, and delayed medical intervention were associated with severe EV71 infection. Cleaning water faucets after hand washing was a protective habit that reduced the risk of complications.

Astrovirus gastroenteritis in hospitalized children of less than 5 years of age in Taiwan, 2009.

BACKGROUND/PURPOSE: Acute gastroenteritis is a common illness in children under 5 years old. Although rotavirus is a leading cause, other viruses including astrovirus are also important, but have been the subject of limited studies. This is a prospective study to investigate astrovirus gastroenteritis in hospitalized children in Taiwan. MATERIAL/METHOD: From January 2009 to December 2009, children below 5 years of age admitted to three hospitals in Taiwan due to acute gastroenteritis were eligible for this study. Stool specimens were sent for the detection of astrovirus by reverse transcriptase polymerase chain reaction; once positive for astrovirus, the sequencing and phylogenetic analysis of each strain was performed. RESULTS: A total of 989 children were enrolled during the study period. The overall positive rate of astrovirus was 1.6%, ranging from 1.03% to 2.26% in different hospitals, while rotavirus accounted for 20.2% of the patients. Six of the 16 children (37.5%) with astroviral infection had documented coinfection with rotavirus. The median age of infection was 28.2 months. The seasonal distribution of astrovirus peaked from April to June. Diarrhea alone (40% vs. 2.1%, p < 0.0001) was significantly more commonly seen than the triad of fever, vomiting and diarrhea (30% vs. 71%, p = 0.0062) in children with astroviral infection alone than in those with rotaviral infection alone. The mean duration of diarrhea was significantly longer in patients with mixed infection than those with astroviral infection alone (6.8 vs. 4.2 days, p = 0.013). Respiratory symptoms were noted in 10 children (62.5%). Serotype HAstV-1 was the most common (68.8%). CONCLUSION: Astrovirus accounted for 1.6% of infections in children under 5 years hospitalized with acute gastroenteritis in Taiwan. Compared with those caused by rotavirus, the incidence of gastroenteritis in hospitalized children caused by astrovirus was low and the disease severity was mild.

[Effects of slopes on nitrogen transport along with runoff from sloping plots on a lateritic red soil amended with sewage sludge].

The effects of different slopes on nitrogen transport along with runoff from sloping plots amended with sewage sludge on a lateritic red soil were studied under simulated rainfall conditions. When the sludge was broadcasted and mixed with surface soils (BM), the MTN (total nitrogen of mixing sample), STN (total nitrogen of settled sample), TPN (total particulate nitrogen), TSN (total suspended nitrogen), TDN (total dissolved nitrogen) and NH4(+) -N concentrations and nitrogen loss amounts in runoff of all treatments were highest at 1 day or 18 days after application. The highest concentrations and the loss amounts of MTN and STN in the slope runoff for the BM treatment increased with slope degree, showing increasing pollution risks to the surface waters. The STN concentration and loss amounts from the 25 degrees plots were 126.1 mg x L(-1) and 1788.6 mg x m(-2), respectively, being 4.6 times and 5.8 times of the corresponding values from the 10 degrees plots, respectively. Then the concentrations and the loss amounts of nitrogen (except NO3(-) -N) from the BM plots diminished rapidly first and then tended to be stable with dwindling differences between the slopes. The loss of MTN and STN in early runoff (1 day and 18 days) accounted for 68.6% -73.4% and 62.3% -66.7% of the cumulative loss amounts during the experimental period for all the broadcasted treatments. Runoff loss coefficients of MTN increased in the order of 20 degrees > 25 degrees > 15 degrees > 10 degrees. Nitrogen was largely lost in dissolved species while large portion of NH4(+) -N was lost with particulates.

Epidemiology and clinical peculiarities of norovirus and rotavirus infection in hospitalized young children with acute diarrhea in Taiwan, 2009.

BACKGROUND/PURPOSE: Acute diarrhea is one of the most common morbidities in pediatrics worldwide. We conducted a study to investigate the incidence of norovirus in young children hospitalized with acute diarrhea in Taiwan and its clinical peculiarity compared with rotavirus gastroenteritis. METHODS: Between January and December, 2009, patients younger than 5 years and admitted to hospital with acute diarrhea were randomly selected; and their stool samples were collected and tested for presence of rotavirus and norovirus by enzyme immunoassay and reverse transcription-polymerase chain reaction, respectively. The clinical manifestations and laboratory findings of the enrolled patients were analyzed. RESULTS: A total of 989 cases were enrolled with a mean age of 21.6 ± 13.7 months and a male proportion of 56.0%. Rotavirus and norovirus was detected in 20.2% and 14.6% of all patients, respectively. Genogroup II was the predominant strain of norovirus (80.6%). Children aged 6-36 months accounted for the majority of patients positive for rotavirus and norovirus (73.0% and 81.3%, respectively). The incidences of norovirus and rotavirus infection were higher during winter and early spring. Most patients with rotavirus and norovirus diarrhea experienced vomiting (74.9%vs. 74.8%, respectively) and fever (94.7%vs. 71.3%, respectively). CONCLUSION: Most young diarrheal patients presenting with vomiting were likely to have norovirus or rotavirus infection. Patients with norovirus diarrhea experienced an absence of, or low-grade fever and longer duration of vomiting compared with those positive for rotavirus infection. A family history of current gastroenteritis may suggest the possibility of norovirus infection.

Transcatheter occlusion of the isolated scimitar vein anomaly camouflaged under dual pulmonary venous drainage of the right lung by the Amplatzer Ductal Occluder.

A 3.5-year-old girl, who presented recurrent pneumonia without discernible cyanosis or desaturation due to isolated scimitar vein anomaly camouflaged by dual pulmonary venous drainage of the right lung, underwent anterograde transvenous occlusion for this abnormal vessel successfully by the Amplatzer Ductal Occluder at 4 years old.

Children's deep neck infections in central Taiwan.

BACKGROUND: Deep neck infection (DNI) is an uncommon but severe disease in children. Few study have been reported detailing pediatric deep neck infection. METHODS: A retrospective analysis of 52 cases of deep neck infection at Changhua Christian Hospital over a 10-year period was performed. RESULTS: Thirty-two boys and 20 girls ranged from one month to eighteen years old were collected in the study. The most remarked symptom was neck swelling (83%). There were 48% (23/48) in right side. Two cases (3.8%) yielded a positive blood culture and fifty-three (53%) cases had positive pus culture. The most common pathogen was Streptococcus viridians (33%). There were two cases oxacillin-resistant Staphylococus aureus. CONCLUSIONS: Once a patient exhibited neck swelling, neck pain, fever and signs of airway obstruction, deep neck infection should be considered. When one was highly suspected to have deep neck infection, computed tomography of neck should be arranged for differential diagnosis.

Clinical course of children of human immunodeficiency virus-infected mothers in Taiwan.

The objective of this study was to describe the clinical courses of infants born to human immunodeficiency virus (HIV)-infected mothers in Taiwan. Eleven children, including 1 set of twins, born to 10 HIV-infected mothers were included in the study. HIV antibodies were assayed and HIV-1 polymerase chain reaction (HIV-PCR) or virus cultures were performed; HIV infection was established when there were at least 2 separate positive results of HIV-PCR or culture. Three sets of primers detecting LTR-gag, pol and env were used. The viral load of HIV RNA was measured and used as an indicator of the treatment response. Two of the children were HIV-infected and received combination therapy, including 2 kinds of nucleoside analogue reverse transcriptase inhibitors plus 1 protease inhibitor. Neither of these children exhibited HIV-related symptoms or signs during the study period. Both mothers of the infected children were Taiwanese and their HIV infection status was not known during pregnancy. In contrast, HIV infection was found in early pregnancy in the 4 women living in Taiwan who were from other countries, all of whom received prophylactic therapy. The other 4 mothers who did not transmit the infection to the infant were Taiwanese, 3 of whom were known to be HIV-seropositive during pregnancy. Based on these results, the vertical rate of transmission was 18% (2/11). Early detection of HIV-infected pregnancy is vital to reduce the incidence of HIV-infected births.

High prevalence of antimicrobial resistance in rapidly growing mycobacteria in Taiwan.

An increasing number of clinical isolations of rapidly growing mycobacteria (RGM) at the National Taiwan University Hospital were noted from 1992 to 2001. Broth microdilution MICs of 15 antimicrobial agents were determined for 200 clinical isolates of RGM, including the Mycobacterium fortuitum group (69 isolates), M. chelonae (39 isolates), and M. abscessus (92 isolates). Our results showed that the resistance rates of these isolates to the currently available agents were remarkably high. Amikacin was active against nearly all RGM isolates. Clarithromycin was usually active against M. abscessus (79% susceptibility) and the M. fortuitum group (65% susceptibility). The majority of M. fortuitum group isolates were susceptible to ciprofloxacin (62%) and imipenem (61%). The susceptibilities to other conventional anti-RGM agents of these isolates were poor but differed markedly by species. The newer fluoroquinolones (levofloxacin, moxifloxacin, and gatifloxacin) and meropenem showed better in vitro activities against the M. fortuitum group isolates than against the other two species of RGM. Linezolid had fairly good activity against these RGM isolates, particularly against M. chelonae isolates (82% susceptible). Telithromycin had poor activity against these RGM isolates (the MICs at which 50% of the isolates tested are inhibited [MIC(50)s] were 32 to 64 microg/ml, and the MIC(90)s were >64 microg/ml).

Clinical manifestations of parainfluenza infection in children.

Parainfluenza viruses are major pathogens causing respiratory illness, manifesting from mild upper respiratory tract infection to bronchiolitis and pneumonia. This retrospective study aimed at providing clinical and epidemiologic data addressing the parainfluenza virus infection in Taiwan. A total of 39 patients were enrolled in this study from March 1999 to December 2000. Infants and young children were the major susceptible population, with 87.2% of them younger than 3 years. No seasonal trend was noted for parainfluenza type 1 and type 2 infections. One clustering of parainfluenza virus type 3 infections occurred in late spring of 2000 based on collected results. Parainfluenza type 1 viral isolates accounted for all of the cases of croup. Most isolates of parainfluenza virus type 3 were associated with upper and/or lower respiratory tract infections. A substantial proportion of the patients had skin involvement; the identification of one case of possible parainfluenza virus-related erythema multiforme is particularly interesting, especially because the chances of a causal relation between viral infection and skin symptoms are formerly thought to be slight. The identification of parainfluenza virus in illnesses classically considered to be due to other viruses is intriguing and may have important implications in the management of childhood illness clinically.