[Efficacy and initial clinical evaluation of optical genome mapping in the diagnosis of structural variations].

To investigate the efficacy and value of optical genome mapping (OGM) in detecting chromosomal structural variations. In a clinical study about high-precision analysis of genomic structural variation for complex genetic diseases, a retrospective study was performed on the cases with karyotyping at the department of Obstetrics and Gynecology, and Endocrinology of Peking Union Medical College Hospital from January to December 2021. Ten cases with abnormal karyotype was detected by OGM. Partial cases were verified by fluorescence in situ hybridization (FISH), SNP array or CNV-seq. Results of ten cases, nine were detected with abnormality by OGM, including unbalanced chromosomal rearrangements (n=3), translocation (n=5) and paracentric inversion (n=1), and the results were in concordance with other standard assays. However, one case with breakpoint and reconnected at centromere has not been detected. In conclusion, ten samples were comprehensively analyzed by karyotyping, FISH, SNP array or CNV-seq, and OGM, and results demonstrated that optical genome mapping as a new technology can not only detect unbalanced rearrangements such as copy number variants as well as balanced translocations and inversions, but more importantly, it can refine breakpoints and orientation of duplicated segments or insertions. So it can contribute to the diagnosis of genetic diseases and prevent birth defect. However, the current technology is not yet capable of detecting breakpoints of balanced structural variations lying within unmapped regions.

End of life care for the most common women cancers in Taiwan.

OBJECTIVES: Women with terminal cancer are assumed to choose hospice care over aggressive treatment at the end of life. With new chemotherapy and target therapy options, it becomes more difficult to decide between hospice care and aggressive management. It is also crucial to consider the cost increases leading to severe financial burdens on healthcare systems. To better understand treatment options at the individual level, this study set out to describe trends in end-of-life care for the four leading cancers in women in Taiwan. STUDY DESIGN: This was a population-based retrospective cohort study. METHODS: The data source was obtained between January 1, 2000, and December 31, 2013, from Taiwan's National Health Insurance Research Database. We identified 98,575 women with a diagnosis of breast (18,596), colorectal (23,734), liver and biliary (28,795) or lung (27,450) cancer who had died during the study period. Hospital data for services provided in the last 6 months of life, including hospice services and aggressive managements (chemotherapy, frequent hospitalisation, emergency room [ER] visits, intensive care unit [ICU] admission and endotracheal intubation), were collected. RESULTS: Hospice utilisation increased over the study period, with 25.85%, 25.34%, 21.23% and 26.55% of female patients with breast, colorectal, liver and biliary, and lung cancer receiving hospice care, respectively. However, the number of women undergoing aggressive treatments in the last 6 months of life remained high, with the breast cancer group having the highest chemotherapy rate, the colorectal cancer group having frequent hospitalisation and the liver and biliary cancer group having frequent ER visits and ICU admissions. CONCLUSIONS: Increasing hospice utilisation among women with the four most common cancers in Taiwan indicates that hospice services have gradually become well accepted over the past 13 years; however, the real focus is on the ineffective treatment preceding hospice care, and late referral was also a notable problem.

Elevated hydrostatic pressure enhances the motility and enlarges the size of the lung cancer cells through aquaporin upregulation mediated by caveolin-1 and ERK1/2 signaling.

The mechanical characteristics presented in cancer microenvironment are known to have pivotal roles in cancer metastasis, which accounts for the leading cause of death from malignant tumors. However, while a uniformly distributed high interstitial fluid pressure (IFP) is a common feature in solid tumors, the effects of high IFP on the motility and invasiveness of cancer cells remain obscure. Using cell-culture devices that simulated increased IFP conditions by applying hydrostatic pressure (HP) ranging from 0 to 20 mm Hg to the cells, we found that the elevated HPs increased the migration speeds, invasiveness, cell volume, filopodial number and aquaporin-1 (AQP1), Snail and vinculin expression levels, as well as phosphorylation of caveolin-1 and extracellular signal-regulated kinase1/2 (ERK1/2), in the lung cancer cells CL1-5 and A549. The increases of migration speed and cell volume correlated temporally with the increase of AQP1 expression. The elevated HP-induced migration acceleration was hindered by AQP1 knockdown using small interfering RNA (siRNA) transfection. Inhibition of ERK1/2 phosphorylation using the mitogen-activated protein kinase kinase inhibitor PD98059 abrogated the elevated HP-induced AQP1 upregulation and migration acceleration in the cancer cells. Caveolin-1 knockdown by siRNA transfection attenuated the HP-induced, ERK1/2-depedent AQP1 upregulation and migration acceleration. Further biochemical studies revealed that the caveolin-1 activation-driven ERK1/2 signaling is mediated by Akt1/2 phosphorylation. By contrast, the elevated HPs had negligible effects on the migration speed and volume of normal bronchial epithelial cells. These results disclose a novel mechanism relating high IFP to the invasiveness of cancer cells and highlight potential targets to impede cancer spreading.

Control of vancomycin-resistant enterococci in a hospital: a five-year experience in a Taiwanese teaching hospital.

In order to prevent transmission of hospital-acquired vancomycin-resistant enterococci (VRE), the infection control team (ICT) of the National Taiwan University Hospital (NTUH) introduced practical guidelines from January 1997 to June 2000. All patients at NTUH found to be infected or colonized with VRE were placed in strict contact and cohort isolation. Surveillance cultures were obtained from other patients in close proximity in order to determine any spread of VRE. If identified, these patients were also placed in contact and cohort isolation, and their isolates were subjected to antimicrobial susceptibility testing and molecular typing by pulsed-field gel electrophoresis. During this period, 20 patients were found to have VRE. Based on typing results, there were three occasions where the same VRE strain had spread between index patients and roommates or patients staying in neighbouring rooms. No further spread occurred after applying strict contact isolation for these patients. The hospital-acquired VRE infection rate was around 0.03 to 0.09 per 1000 discharges during the intervention period. After July 2000, however, members of the ICT did not actively monitor or implement any interventions to control VRE. The rate then increased to 0.20 per 1000 discharges in 2001. This study suggests that interventions for the control of VRE, based on the guidelines from the Hospital Infection Control Practice Advisory Committee, are effective for control of VRE spread. Failure to adhere to these guidelines may result in an increase in hospital-acquired VRE.

Healthcare-associated outbreak due to pan-drug resistant Acinetobacter baumannii in a surgical intensive care unit.

Acinetobacter baumannii is ubiquitous and has recently become one of the most important healthcare-associated (HA) pathogens in hospitals. Infection caused by this organism often leads to significant morbidity and mortality. Outbreaks of pan-drug resistant Acinetobacter baumannii (PDRAB) have rarely been reported. During a two-month period, an outbreak of PDRAB colonization and infection affecting 7 patients occurred in our surgical intensive care unit (SICU). The colonized sites were respiratory tract (N = 7) and central venous catheter (N = 2). One of the patients had a surgical wound infection. Extensive environmental contamination was identified, including sites such as bed rails, bedside tables, surface of ventilators and infusion pump, water for nasogastric feeding and ventilator rinsing and sinks. All of the isolates were analysed by pulsed-field gel electrophoresis (PFGE) and showed an identical pattern. After use of strict cohort nursing, hand hygiene environmental cleaning, and replacement of a dysfunctional high-efficiency particulate air filter (HEPA), the outbreak was controlled.

Nosocomial Exophiala jeanselmei pseudoinfection after sonography-guided aspiration of thoracic lesions.

BACKGROUND AND PURPOSE: During the period from August 1994 to October 1998, a total of 19 isolates of Exophiala jeanselmei were recovered from 17 patients with various underlying thoracic diseases treated at National Taiwan University Hospital. The purpose of this study was to describe the clinical characteristics of these patients and to determine the microbiologic relatedness of the E. jeanselmei. METHODS: Of the 19 isolates, 11 from nine patients were preserved and were identified based on their biotypes as determined by the API ID32C System, their cellular fatty acid profiles by gas-liquid chromatography, their antibiotypes to five antifungal agents by the E-test, and their random amplified polymorphic DNA (RAPD) patterns by arbitrarily primed PCR. Extensive environmental surveillance cultures and cultures from the skin of eight patients and hands of one physician were also performed. RESULTS: One of the 17 patients had E. jeanselmei isolated from cutaneous phaeohyphomycosis (3 isolates), and the other 16 patients had isolations from pleural effusion specimens (15 isolates) or lung mass (1 isolate) following sonography-guided aspiration. The latter 16 patients had no clinical or pathologic evidence of fungal infection. Isolates (clone 1) from aspirated specimens had identical biotypes, antibiotypes, and RAPD patterns, which were different from those of the three isolates (clone 2) from the patient with a cutaneous lesion. All specimens from environmental sources, patients' skin, and the hands of the physician were negative for E. jeanselmei. CONCLUSION: This series of patients demonstrates the difficulty in identifying the sources of a nosocomial pseudoinfection due to this slow-growing microorganism when isolated from pleural effusion specimens.

Persistent bacteraemia caused by a single clone of Burkholderia cepacia with unusual phenotype.

We report a case of persistent bacteraemia caused by a single clone of Burkholderia cepacia with unusual characteristics. Six isolates of B. cepacia were recovered from a patient with acute myeloid leukaemia and chemotherapy-induced neutropenia within a 3-week period. All six isolates were initially incompletely identified as B. cepacia with the API 20NE system. The further use of cellular fatty acid analysis and PCR-restriction fragment length polymorphism of the 16S rDNA confirmed the identification. These isolates also displayed an identical but unusual antibiotype. The identical cellular fatty acid profiles and genomic typing generated by random amplified polymorphic DNA identified these isolates as derivatives of a single strain.

A hospital-acquired outbreak of methicillin-resistant Staphylococcus aureus infection initiated by a surgeon carrier.

Methicillin-resistant Staphylococcus aureus (MRSA) has become an important hospital-acquired pathogen, infection with which often leads to major morbidity and mortality. The principal mode of transmission for MRSA is transfer of the organism from a carrier or infected patient to uninfected patients by the hands or clothing of staff. From January 16 1997 to April 2 1997, five patients who had undergone open-heart surgery in a hospital located in northern Taiwan, developed surgical wound infections and mediastinitis caused by MRSA. All patients were hospitalized in two adjacent surgical intensive care units (ICUs) following their respective operations. Consequently, the hospital's infection control team commenced investigation of the outbreak. Pulsed-field gel electrophoresis (PFGE) has been shown to be a good technique for epidemiological typing. By analysing cultures taken from staff by PFGE, it was demonstrated that this outbreak was most likely to be initiated by a surgeon with MRSA carriage. After elimination of the carrier state using topical mupirocin treatment, the outbreak was controlled without further incident.

Quinupristin-dalfopristin resistance among gram-positive bacteria in Taiwan.

To understand quinupristin-dalfopristin resistance among clinical isolates of gram-positive bacteria in Taiwan, where this agent is not yet available for clinical use, we evaluated 1,287 nonduplicate isolates recovered from January 1996 to December 1999 for in vitro susceptibility to quinupristin-dalfopristin and other newer antimicrobial agents. All methicillin-susceptible Staphylococcus aureus (MSSA) isolates were susceptible to quinupristin-dalfopristin. High rates of nonsusceptibility to quinupristin-dalfopristin (MICs, >/=2 microg/ml) were demonstrated for the following organisms: methicillin-resistant S. aureus (MRSA) (31%), coagulase-negative staphylococci (CoNS) (16%), Streptococcus pneumoniae (8%), viridans group streptococci (51%), vancomycin-susceptible enterococci (85%), vancomycin-resistant Enterococcus faecalis (100%), vancomycin-resistant Enterococcus faecium (66%), Leuconostoc spp. (100%), Lactobacillus spp. (50%), and Pediococcus spp. (87%). All isolates of MSSA, MRSA, S. pneumoniae, and viridans group streptococci were susceptible to vancomycin and teicoplanin. The rates of nonsusceptibility to vancomycin and teicoplanin were 5 and 7%, respectively, for CoNS, ranging from 12 and 18% for S. simulans to 0 and 0% for S. cohnii and S. auricularis. Moxifloxacin and trovafloxacin had good activities against these isolates except for ciprofloxacin-resistant vancomycin-resistant enterococci and methicillin-resistant staphylococci. In Taiwan, virginiamycin has been used in animal husbandry for more than 20 years, which may contribute to the high rates of quinupristin-dalfopristin resistance.

Lack of androgen receptor transcriptional activity in human keratinocytes.

Since detection of androgen receptor (AR) expression in keratinocytes by immunostaining is controversial, we investigated whether keratinocytes can act as androgen target cells using transient transfection assays. Chloramphenicol acetyltransferase (CAT) assays for the endogenous AR transcriptional activity in HaCaT keratinocytes indicated that DHT (10(-9)-10(-8) M) can induce less than 1.5-fold of mouse mammary tumor virus CAT, which is quite low, compared with 38-fold induction by 10(-7) M 1,25-dihydroxyvitamin D(3) of P450cc24-CAT. Furthermore, this low DHT-mediated induction could not be enhanced by the AR co-activators, ARA70 or ARA55. Western blotting analysis indicated that HaCaT and normal keratinocytes do not express AR protein. Transfection of exogenous AR into HaCaT keratinocytes, however, could install AR transcriptional activity, suggesting that HaCaT keratinocytes have all the necessary accessory factors for AR transcription activity. In conclusion, keratinocytes are unlikely to be target cells for androgen.

Application of pulsed-field gel electrophoresis to the investigation of a nosocomial outbreak of Vibrio parahaemolyticus.

The method of pulsed-field gel electrophoresis (PFGE) for Vibrio parahaemolyticus was first published in 1996. Since then, its application has been seldom reported in the literature. A food poisoning outbreak due to V. parahaemolyticus occurred in five wards of a hospital was investigated using this method. Twenty-five patients were involved and all of them had eaten food supplied by the hospital. Of the 15 cases whose stools were available for culture, only four cultures yielded V. parahaemolyticus. All four isolates were serotype K6 and were indistinguishable or closely related to each other based on PFGE patterns. Two isolates from food were recovered and they presented different characterizations from the patient isolates in both serotype and PFGE pattern. Successful typing by PFGE to identify the outbreak strain and differentiate V. parahaemolyticus strains between patient and food isolates in this study suggests the usefulness of PFGE for V. parahaemolyticus, the leading cause of food poisoning in Taiwan.

High protease activity of Chryseobacterium indologenes isolates associated with invasive infection.

In order to understand virulence factors of Chryseobacterium indologenes isolates associated with invasive infection, enzymatic activities and cellular fatty acid profiles of 42 isolates recovered at National Taiwan University Hospital from January 1994 to December 1996 were studied. Among them, 12 blood isolates were considered as invasive and 30 (recovered from urine, sputa, infected burn wounds, and catheter tips) were noninvasive. All isolates showed strong activities of alkaline phosphatase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, and N-acetyl-beta-glucosaminidase, and had no activities for alpha-galactosidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, and alpha-fucosidase. The activities of other enzymes were variable. Thirty-two isolates (76%) had varying degrees of protease activity. Two profiles (profiles I and II) of cellular fatty acids of the isolates were found and profile I predominated. There was no significant difference of distribution of cellular fatty acid profiles and activities of enzymes between invasive and noninvasive isolates, except protease activity which was significantly higher in invasive isolates than that in noninvasive isolates. Protease activity may play an important role in virulence on invasive infections caused by C. indologenes.

[Three dimensional Finite Element Analysis of the Effects of Posts and Cores with Different Angles]

OBJECTIVE:The test is an analogue of the conditions that cast post and core can be made with different angles to get the core incline lingually or facially in clinic, so that the arrangement of the anterior teeth may be improved.METHODS:Analyse the stress distribution and concentration in the dentin of posts and cores with different angles by Ansys three dimensional finite element stress analysis method.RESULTS:The stress in root was positively related to the angles between the load and the axis of the tooth, the stress distributoin in dentin was related to the direction of the core but not related to the angles. The closer the distance from the load to cemento enamel junction,the greater the effect of the load on the stress in dentin.CONCLUSION:When the cast post and core with angle was used to rectify the facially malpositional maxillary anterior teeth, the core should not be inclined lingually excessively. It is important to assure the thickness of the facial and lingual canal wall during tooth preparing and to adjust the occlusion so that the root fracture can be avoided.

Emergence of vancomycin-resistant enterococci at a university hospital in Taiwan: persistence of multiple species and multiple clones.

OBJECTIVES: To describe the epidemiology of vancomycin-resistant enterococci (VRE) in a university hospital in Taipei, Taiwan. DESIGN: Retrospective review over a 27-month period, from March 1996 to May 1998. SETTING: A tertiary-care teaching hospital in Taiwan. PARTICIPANTS: Patients with VRE isolated from any body site. METHODS: Patients were identified through hospital microbiology and infection control records. Patient charts were reviewed for clinical and epidemiology data, including age, gender, previous hospital admissions, underlying diseases, types of infection, and recent antibiotic use. VRE isolates were characterized by their typical biochemical reactions, cellular fatty acid profiles, and the presence of van genes. Antibiotypes using the E-test and randomly amplified polymorphic DNA (RAPD) patterns of these isolates were used to determine the clonality. RESULTS: Twenty-five isolates of VRE recovered from 12 patients were identified. One patient with a perianal abscess had 12 isolates of VRE (4 Enterococcus faecalis, 7 Enterococcus faecium, and 1 Enterococcus casseliflavus) recovered from perianal lesions. Among 3 patients who were hospitalized in the same room, 1 had a community-acquired cellulitis over the left leg caused by E. faecalis, and the other 2 patients both had anal colonization with 2 isolates of E. faecalis. The other 8 patients had 1 E. faecalis isolate each from various clinical specimens. All isolates possessed vanA resistance phenotype and vanA genes. Different antibiotypes and RAPD patterns of the isolates from different patients excluded the possibility of nosocomial spread at the hospital. CONCLUSIONS: Multiple species of VRE (E. faecalis, E. faecium, and E. casseliflavus) and multiple clones of E. faecium could colonize or infect hospitalized patients. In addition, clones of VRE can persist long-term in patients' lower gastrointestinal tracts. These results extend our knowledge of the coexistence and the persistence of multiple species and multiple clones of VRE in hospitalized patients.

Longitudinal analysis of methicillin-resistant Staphylococcus aureus isolates at a teaching hospital in Taiwan.

In Taiwan, the frequency of nosocomial infections caused by methicillin-resistant Staphylococcus aureus (MRSA) has increased rapidly during the past 10 years. To investigate the epidemiology of MRSA infections, a total of 140 MRSA isolates collected at National Taiwan University Hospital from 1992 to 1996 were characterized by pulsed-field gel electrophoresis (PFGE) profiles and antibiotypes, as determined with the disk diffusion method. Among these isolates, six PFGE types (with 20 subtypes) and six antibiotypes were identified. Antibiotyping proved to be a poor method of epidemiologic analysis, because almost all of the MRSA isolates analyzed shared a very similar multidrug-resistant antibiotype. Most MRSA infections and colonizations in this hospital were due to the spread of strains belonging to three major PFGE types (A, B, and C). However, the major type changed in different years with types A, B, and C being predominant in 1992 through 1993, 1994 through 1995, and 1996, respectively. The three major PFGE types spread easily throughout the hospital wards, presumably carried by health care workers and environmental contamination. Our results demonstrate that there was a dominant strain spreading in our hospital each year and the dominant strain may shift in different years.

Transgenic mouse model of AA amyloidosis.

AA amyloidosis can be induced in mice experimentally through injection of certain chemical or biological compounds. However, the usefulness of this approach is limited by its dependence on exogenous inflammatory agents that stimulate cytokines to increase the synthesis of precursor serum amyloid A (SAA) protein and the transitory nature of the pathological fibrillar deposits. We now report that transgenic mice carrying the human interleukin 6 gene under the control of the metallothionein-I promoter had markedly increased concentrations of SAA and developed amyloid in the spleen, liver, and kidneys by 3 months of age. At the time of death about 6 months later, organs obtained from these animals had extensive amyloid deposits. This disease process was apparent radiographically using small-animal computer axial tomography and magnetic resonance imaging equipment. The AA nature of the amyloid was evidenced immunohistochemically and was unequivocally established by sequence analysis of protein extracted from the fibrils. The availability of this unique in vivo experimental model of AA amyloidosis provides the means to assess the therapeutic efficacy of agents designed to reduce or prevent the fibrillar deposits found in AA and other types of amyloid-associated disease.

Dissemination of two methicillin-resistant Staphylococcus aureus clones exhibiting negative staphylase reactions in intensive care units.

From December 1997 to March 1998, 25 methicillin-resistant Staphylococcus aureus (MRSA) isolates exhibiting negative Staphylase (Oxoid Ltd., Basingstoke, England) reactions were identified from various clinical specimens from 13 patients in six intensive care units (ICUs) or in wards following a stay in an ICU at the National Taiwan University Hospital. The characteristics of these isolates have not been previously noted in other MRSA isolates from this hospital. Colonies of all these isolates were grown on Trypticase soy agar supplemented with 5% sheep blood and were nonhemolytic and unpigmented. Seven isolates, initially reported as Staphylococcus haemolyticus (5 isolates) and Staphylococcus epidermidis (2 isolates) by the routine identification scheme and with the Vitek GPI system (bioMerieux Vitek, Inc., Hazelwood, Mo.), were subsequently identified as S. aureus by positive tube coagulase tests, standard biochemical reactions, and characteristic cellular fatty acid chromatograms. The antibiotypes obtained by the E test, coagulase types, restriction fragment length polymorphism profiles of the staphylococcal coagulase gene, and random amplified polymorphic DNA patterns generated by arbitrarily primed PCR of the isolates disclosed that two major clones disseminated in the ICUs. Clone 1 (16 isolates) was resistant to clindamycin and was susceptible to trimethoprim-sulfamethoxazole (TMP-SMZ) and was coagulase type II. Clone 2 (eight isolates) was resistant to clindamycin and TMP-SMZ and was coagulase type IV. These two epidemic clones from ICUs are unique and underline the need for caution in identifying MRSA strains with colonial morphologies not of the typical type and with negative Staphylase reactions.

Roles of testosterone in the growth of keratinocytes through bald frontal dermal papilla cells.

A coculture of dermal papilla cells (DPCs) from the bald frontal scalp of stumptailed macaques with keratinocytes derived from human neonatal foreskin revealed that testosterone inhibited keratinocyte proliferation, and that the antiandrogen RU58841 abolished this response. This testosterone-induced keratinocyte growth inhibition was not observed when either type of cells was cultured alone. We also examined conditioned media from the coculture system and demonstrated the identical testosterone-induced growth inhibition on keratinocytes, and this inhibitory effect was conditioned media concentration-dependent. These results suggested that the testosterone-mediated suppression on keratinocyte proliferation might proceed through some diffusible growth mediators in conditioned media. Differential display reverse transcriptase polymerase chain reaction allowed us to isolate several genes from frontal DPCs that can be either suppressed or induced by testosterone. Supervillin, a membrane-associated, F-actin-binding protein, was identified as one of the testosterone downregulated genes in frontal DPCs. Further characterization of these testosterone-target genes may reveal the mechanism by which testosterone inhibits the growth of follicular cells in androgenetic alopecia.

Evaluation of RU58841 as an anti-androgen in prostate PC3 cells and a topical anti-alopecia agent in the bald scalp of stumptailed macaques.

The effect of androgen receptor transcriptional activation by RU58841, a nonsteroidal anti-androgen, was studied in the human prostate cancer PC3 cell line by cotransfection with wild-type androgen receptor (wt AR) and an androgen-responsive reporter (MMTV-ARE-CAT) construct. Anti-and rogens, hydroxyflutamide, and Casodex, and the antiestrogen, genistein, were studied in parallel for comparison with RU58841. The wt AR was activated only by the androgen dihydrotestosterone (DHT). Neither the anti-androgens nor antiestrogen can enhance AR transcriptional activity at 10(-11)-10(-7)M in PC3 cells. Hydroxyflutamide, RU58841, and Casodex, but not genistein, displayed competitively suppressive effects on DHT activation of wt AR. The potency of RU58841 was comparable to that of hydroxyflutamide. From this result, topical application of RU58841, which is considered to be a potential therapy for skin diseases, may induce systemic side effects. However, RU58841, on topical application, revealed a potent increase in density, thickening, and length of hair in the macaque model of androgenetic alopecia, whereas no systemic effects were detected. Together our results suggest that RU58841 may have potent antagonism to the wt AR and could be considered as a topically applied active anti-androgen for the treatment of androgen-dependent skin disorders, such as acne, androgenetic alopecia, and hirsutism.