Coping strategies of intensive care units nurses in alarm management: a qualitative research study.

BACKGROUND: Intensive care units are critical environments where various alarm systems play a pivotal role in patient monitoring and safety. Alarm fatigue can lead to slower response times and missed alarms, compromising patient safety and increasing stress and burnout among intensive care unit nurses. Understanding how intensive care unit nurses respond to and manage these alarms is crucial in evaluating their impact on patient care and nursing well-being. METHODS: This descriptive qualitative study explored the experiences of intensive care unit nurses in alarm management. Conducted in the medical and surgical intensive care units of a Northern Taiwan medical center, the study involved 15 nurses. Semi-structured interviews were utilized to investigate the working experiences of ICU nurses in alarm management and to identify their coping strategies for dealing with the constant inundation of medical device alarms. The interviews were transcribed, and content analysis was applied to identify key themes in the responses. RESULTS: The study revealed five main themes in intensive care unit nurses' strategies for managing alarms: (1) Mastering alarm signals and acting; (2) Team monitoring for life preservation; (3) Enhancing senses and distinguishing carefully; (4) Learning from the lessons of incidents for vigilant reflection; and (5) Detach alarms' influence on daily life. These coping strategies are effective in alarm management, safeguarding patients' lives, enhancing the serenity of the clinical environment, and mitigating the physical and mental exhaustion caused by alarm fatigue. CONCLUSIONS: Intensive Care Unit nurses develop various coping strategies to manage medical device alarms, based on their experience. These strategies are crucial in maintaining patient safety and reducing nurse alarm fatigue. They can also be used for nursing education and clinical training.

The Effect of the Knowledge, Skills, and Attitudes from Nurse Training Using In Situ Simulation in an Intensive Care Unit.

BACKGROUND: In situ simulation is the practice of using simulated scenarios to improve skill implementation, train critical thinking and problem-solving abilities, and enhance self-efficacy. This study aimed to enhance nursing knowledge, skills, and attitudes toward clinical work by applying in situ simulation training to improve the healthcare of critically ill patients. METHODS: This study was conducted from a medical center in northern Taiwan and included 86 trainees who received intensive care training courses from 1 June 2017 to 31 May 2019. The self-report knowledge assessment, empathetic self-efficacy scale, skill assessment, and attitudes of instructors before and after training were collected. The statistical analysis used the Wilcoxon test for knowledge and attitudes, and chi-square tests were used for skills to evaluate the learning effect. RESULTS: The results showed a statistically significant improvement in knowledge, skills, attitudes, and empathy in nursing care. CONCLUSIONS: In situ simulation learning can be an accepted method for nursing skills in the intensive care unit. Through this study, we understood that the in situ simulation method was beneficial to nurses' care and care thinking processes. It is worth developing and evaluating integrated simulation education to enhance learning, change behavior, and promote holistic care in the nursing field.

Reduction in the Incidence Density of Pressure Injuries in Intensive Care Units after Advance Preventive Protocols.

(1) Background: Patients who are critically ill or undergo major surgery are admitted to intensive care units (ICUs). Prolonged immobilization is the most likely cause of pressure injuries (PrIs) in the ICU. Previous studies of Western populations found that effective protocols could reduce the incidence of PrIs, and the efficacy of systemic targeted intervention protocols in preventing PrIs in the Chinese population needs to be surveyed. (2) Methods: We reviewed cases of PrIs in the ICUs of Taipei Veterans General Hospital from 2014 to 2019. The ICU nurses at the hospital began to implement targeted interventions in January 2017. The incidence density of PrIs was calculated by dividing the number of PrIs by person days of hospitalizations in the pre-bundle (2014-2016) and post-bundle (2017-2019) stages. Poisson regression was performed to compare the trend of incidence densities. (3) Results: The incidence density of PrIs was 9.37/1000 person days during the pre-bundle stage and 1.85/1000 person days during the post-bundle stage (p < 0.001). The relative risk (RR) was 0.197 (95% confidence interval: 0.149-0.26). The incidence densities of iatrogenic PrIs and non-iatrogenic PrIs decreased as the RRs decreased. (4) Conclusions: Targeted interventions could significantly reduce the incidence of PrIs. Healthcare providers must follow the bundle care protocol for PrI prevention to improve the quality of healthcare and promote patient health.

[The Impact of Critical Care Transition Programs (CCTPs) on Relocation Stress in Patients and Patient Families in the Intensive Care Unit].

The intensive care unit (ICU) is designed to care for patients with high disease severity who require critical care and close monitoring. Patients in the ICU may be transferred to the general ward for further treatment following recovery from the acute phase. Transferring from the ICU to the general ward after acute phase recovery is a stressful event that may stress both the patient and their family, potentially resulting in relocation stress syndrome (RSS). RSS has been found to be closely related to unplanned ICU readmissions, prolonged hospitalization, and adverse events, affecting recovery to health and family peace of mind. Furthermore, RSS may result in ineffective disease coping, feelings of uncertainty, and poor treatment response, leading to prolonged hospitalization, reduced trust in medical staff, and decreased happiness and quality of life. In recent years, the nursing profession has attached increasing importance to holistic health care. This has encouraged critical care teams to map out customized relocation plans for patients who are about to be transferred from the ICU that use standardized evaluation tools for transfers and elicit the situation and needs from patients or their family. Through the setting of care goals, shared decision making, and cross-unit support, enhanced communications among the medical team facilitate transition preparation and improve the quality and effect of intensive care.

Validation of the prophylactic efficacy of urea-based creams on sorafenib-induced hand-foot skin reaction in patients with advanced hepatocellular carcinoma: A randomised experiment study.

BACKGROUND: Hand-foot skin reaction may influence the effectiveness of patients' treatment, patient quality of life, and the economics of health care. An effective prophylactic dermatological cream for preventing sorafenib-induced hand-foot skin reaction (HFSR) is yet to be identified. AIM: The aim of this study is validated the prophylactic efficacy of urea-based creams on sorafenib-induced hand-foot skin reaction in patients with advanced hepatocellular carcinoma. METHODS: This was a randomised double-blind experimental study. A total of 129 patients with advanced HCC were randomly assigned to three groups. The comparison group received best supportive care (BSC), group A received BSC plus a moisturising cream, and group B received BSC plus a 10% urea-based cream. Incidence of HFSR and cutaneous wetness were assessed 3 days before starting sorafenib and each week after starting sorafenib for 8 weeks. RESULTS: No significant difference was observed in the incidence density of sorafenib-induced HFSK (comparison group/A group, p > .05; comparison group/B group, p > .05). Group B reported significantly better cutaneous wetness of hands in the seventh week after starting sorafenib (p < .05) and of feet during the first 6 weeks (p < .05-.001). CONCLUSION: This study found a nut size amount of a 10% urea-based cream applied twice a day can maintain patients' cutaneous wetness in the first 6 weeks after starting sorafenib than moisturising-alone cream. But it cannot reduce the occurrence of HFSR. Thus, the result supports nut-size dose of the 10% urea-based cream three times a day may be an appropriate dose to prevent HFSR. Clinical Trail Registration Number: NCT04568330.

Paradox of trimethylamine-N-oxide, the impact of malnutrition on microbiota-derived metabolites and septic patients.

BACKGROUND: Trimethylamine N-oxide (TMAO) is a microbiota-derived metabolite, which is linked to vascular inflammation and atherosclerosis in cardiovascular (CV) diseases. But its effect in infectious diseases remains unclear. We conducted a single-center prospective study to investigate association of TMAO with in-hospital mortality in septic patients admitted to an intensive care unit (ICU). METHODS: Totally 95 septic, mechanically ventilated patients were enrolled. Blood samples were obtained within 24 h after ICU admission, and plasma TMAO concentrations were determined. Septic patients were grouped into tertiles according to TMAO concentration. The primary outcome was in-hospital death, which further classified as CV and non-CV death. Besides, we also compared the TMAO concentrations of septic patients with 129 non-septic patients who were admitted for elective coronary angiography (CAG). RESULTS: Septic patients had significantly lower plasma TMAO levels than did subjects admitted for CAG (1.0 vs. 3.0 µmol/L, p < 0.001). Septic patients in the lowest TMAO tertile (< 0.4 µmol/L) had poorer nutrition status and were given longer antibiotic courses before ICU admission. Circulating TMAO levels correlated positively with daily energy intake, the albumin and prealbumin concentration. Compared with those in the highest TMAO tertile, septic patients in the lowest TMAO tertile were at greater risk of non-CV death (hazard ratio 2.51, 95% confidence interval 1.21-5.24, p = 0.014). However, TMAO concentration was no longer an independent predictor for non-CV death after adjustment for disease severity and nutritional status. CONCLUSION: Plasma TMAO concentration was inversely associated with non-CV death among extremely ill septic patients, which could be characterized as TMAO paradox. For septic patients, the impact of malnutrition reflected by circulating TMAO levels was greater than its pro-inflammatory nature.

Elevated serum galectin-1 concentrations are associated with increased risks of mortality and acute kidney injury in critically ill patients.

BACKGROUND: Galectin-1 (Gal-1), a member of the ß-galactoside binding protein family, is associated with inflammation and chronic kidney disease. However, the effect of Gal-1 on mortality and acute kidney injury (AKI) in critically-ill patients remain unclear. METHODS: From May 2018 to March 2020, 350 patients admitted to the medical intensive care unit (ICU) of Taipei Veterans General Hospital, a tertiary medical center, were enrolled in this study. Forty-one patients receiving long-term renal replacement therapy were excluded. Serum Gal-1 levels were determined within 24 h of ICU admission. The patients were divided into tertiles according to their serum Gal-1 levels (low, serum Gal-1 < 39 ng/ml; median, 39-70 ng/ml; high, ≥71 ng/ml). All patients were followed for 90 days or until death. RESULTS: Mortality in the ICU and at 90 days was greater among patients with elevated serum Gal-1 levels. In analyses adjusted for the body mass index, malignancy, sepsis, Sequential Organ Failure Assessment (SOFA) score, and serum lactate level, the serum Gal-1 level remained an independent predictor of 90-day mortality [median vs. low: adjusted hazard ratio (aHR) 2.11, 95% confidence interval (CI) 1.24-3.60, p = 0.006; high vs. low: aHR 3.21, 95% CI 1.90-5.42, p < 0.001]. Higher serum Gal-1 levels were also associated with a higher incidence of AKI within 48 h after ICU admission, independent of the SOFA score and renal function (median vs. low: aHR 2.77, 95% CI 1.21-6.34, p = 0.016; high vs. low: aHR 2.88, 95% CI 1.20-6.88, p = 0.017). The results were consistent among different subgroups with high and low Gal-1 levels. CONCLUSION: Serum Gal-1 elevation at the time of ICU admission were associated with an increased risk of mortality at 90 days, and an increased incidence of AKI within 48 h after ICU admission.

[The Relationship Between Life Attitude and Death Coping Self-Efficacy in Intensive Care Nurses].

BACKGROUND: Critical care nurses must often care for patients who are dying and their families. Thus, understanding the self-efficacy and life attitudes of nursing staff in the ICU in response to death is important to the development and provision of relevant education and training. PURPOSE: This study was designed to explore the self-efficacy of ICU nurses in response to death and related predictive factors. METHODS: This was a cross-sectional research study. The subjects were 216 nurses in the adult ICU of a medical center in northern Taiwan. The research tools used included the death coping self-efficacy scale and the life attitude scale. Data were analyzed using Pearson's correlation, t-test, one-way ANOVA, and multiple regression. RESULTS: The results showed: 1. In terms of death coping self-efficacy, the mean score was 112.0 ± 14.3, with the highest scoring subscale, hospice care, earning a mean score of 51.1 ± 6.3. In terms of life attitude, the mean score was 128.9 ± 13.8, with the highest scoring subscale, life autonomy, earning a mean score of 24.0 ± 3.2. 2. Nurses with experiences of withdrawal of life support had better coping efficacy (t = 1.94, p = .05) and those with a graduate degree or above earned a better average life attitude score than those educated to the university / junior college level. 3. Age and ICU seniority were found to correlate positively with grief-related coping skills (r = .241- .315), with the life-attitude subscales of aspiring, life-autonomy, love, and caring showing positive correlations with death coping self-efficacy (r = .138- .482). 4. The predictors found in this study for death coping self-efficacy were age, aspiring, life-autonomy, love, and caring, with a total explained variance of 30.1% (F = 12.78, p < .001). CONCLUSIONS: The results of this study indicate that education level and having hospice care experience are both significant predictors of life attitude in ICU nurses, which is a factor that is known to affect self-efficacy in response to death. Life attitude and hospice care training programs for ICU nurses should be promoted to foster positive life attitudes and thereby enhance self-efficacy in response to death to improve the quality of intensive clinical care.

[Sepsis and Early Warning Score Systems].

Sepsis is a significant cause of morbidity and mortality worldwide. Early diagnosis and management of sepsis is critical to improving patient prognoses. Surviving sepsis campaign guidelines issued in 2016 encourage health institutions to establish a screening system to identify patients who are at risk of sepsis. In 2012, the Royal College of Physicians in the UK began to advocate replacing local and regional scoring systems with the National Early Warning Score (NEWS), which is optimized for the identification of sepsis. Although many hospitals continue to use other scoring systems, all healthcare organizations are being encouraged to adopt a standardized scoring system to better promote patient safety by facilitating rapid diagnoses and screenings and thus, subsequently, improving decision-making by clinical staffs. NEWS plays a very important role in the treatment of sepsis patients. Although research findings related to this scoring system differ somewhat, they provide an important reference for clinical nursing staffs. Intelligent systems are not comprehensive in terms of their capabilities. However, combining human intelligence with system features and further optimizing the system should contribute significantly to the reduction of mortality risk in patients with sepsis.

Lipid paradox in patients with acute myocardial infarction: Potential impact of malnutrition.

BACKGROUND & AIMS: Aggressive lipid reduction is recommended for patients with AMI, but reverse epidemiology, the lipid paradox, has been reported in several clinical studies. The cause of lipid paradox remains uncertain, and nutrition is one possible explanation. In this single-center retrospective study, we investigated the relationships between baseline LDL concentrations and clinical outcomes in patients with AMI, stratified by different nutritional status. METHODS: Totally 409 patients were enrolled for analysis. The Nutritional Risk Index (NRI) was used to estimate the risk of malnutrition. Subjects were grouped into tertiles according to their NRIs. Clinical outcomes were compared among patients with varying NRIs and LDL levels. RESULTS: Patients in the lowest NRI tertile had increased incidences of in-hospital mortality, cardiogenic shock, decompensated heart failure, renal failure, and sepsis. This tertile was also associated with increased long-term mortality during the follow-up period of 832 ± 744 days. Mortality was increased among patients with baseline LDL concentrations ≤70 mg/dL in the lowest NRI tertile (log rank test, p = 0.0257), but not in the high or median tertiles. Moreover, baseline LDL level ≤70 mg/dL was an independent risk factor of all-cause mortality (adjusted hazard ratio = 1.73; 95% confidence interval, 1.01-2.94; p = 0.045) in the lowest NRI tertile. CONCLUSIONS: Lipid paradox was observed in the high-risk of malnutrition population among patients with AMI. Aggressive lipid-lowering therapy is still recommended for patients with AMI and fair nutritional status. However, when treating patients at high risk of malnutrition, the improvement of nutritional status may be more beneficial than strict LDL control.

Aberrant Sensory Gating of the Primary Somatosensory Cortex Contributes to the Motor Circuit Dysfunction in Paroxysmal Kinesigenic Dyskinesia.

Paroxysmal kinesigenic dyskinesia (PKD) is conventionally regarded as a movement disorder (MD) and characterized by episodic hyperkinesia by sudden movements. However, patients of PKD often have sensory aura and respond excellently to antiepileptic agents. PRRT2 mutations, the most common genetic etiology of PKD, could cause epilepsy syndromes as well. Standing in the twilight zone between MDs and epilepsy, the pathogenesis of PKD is unclear. Gamma oscillations arise from the inhibitory interneurons which are crucial in the thalamocortical circuits. The role of synchronized gamma oscillations in sensory gating is an important mechanism of automatic cortical inhibition. The patterns of gamma oscillations have been used to characterize neurophysiological features of many neurological diseases, including epilepsy and MDs. This study was aimed to investigate the features of gamma synchronizations in PKD. In the paired-pulse electrical-stimulation task, we recorded the magnetoencephalographic data with distributed source modeling and time-frequency analysis in 19 patients of newly-diagnosed PKD without receiving pharmacotherapy and 18 healthy controls. In combination with the magnetic resonance imaging, the source of gamma oscillations was localized in the primary somatosensory cortex. Somatosensory evoked fields of PKD patients had a reduced peak frequency (p < 0.001 for the first and the second response) and a prolonged peak latency (the first response p = 0.02, the second response p = 0.002), indicating the synchronization of gamma oscillation is significantly attenuated. The power ratio between two responses was much higher in the PKD group (p = 0.013), indicating the incompetence of activity suppression. Aberrant gamma synchronizations revealed the defective sensory gating of the somatosensory area contributes the pathogenesis of PKD. Our findings documented disinhibited cortical function is a pathomechanism common to PKD and epilepsy, thus rationalized the clinical overlaps of these two diseases and the therapeutic effect of antiepileptic agents for PKD. There is a greater reduction of the peak gamma frequency in PRRT2-related PKD than the non-PRRT PKD group (p = 0.028 for the first response, p = 0.004 for the second response). Loss-of-function PRRT2 mutations could lead to synaptic dysfunction. The disinhibiton change on neurophysiology reflected the impacts of PRRT2 mutations on human neurophysiology.

[Prevention and nursing care of central line-associated bloodstream infections in critically ill patients].

Catheter-related bloodstream infections are associated with significantly increased morbidity, mortality, and expenditures. Such infections are a serious threat to patient safety in the intensive care unit. This review describes the latest protocols related to preventing and treating central venous catheter-associated bloodstream infections in critically ill patients. According to 2011 Center for Disease Control (CDC) guidelines and central line care bundles by the institute for healthcare improvement (IHI), prevention measures for catheter-related bloodstream infections include the following: hand hygiene, maximal barrier precautions insertion, chlorhexidine skin antisepsis, optimal catheter site selection, proper catheter maintenance, insertion site care, and daily review of line necessity, with prompt removal of unnecessary lines. These are important and effective infection prevention measures. Guidelines and care bundles also recommend organizing care modules based on unit characteristics; integrating resources and empirical measures; education and training to promote comprehensive implementation; and auditing and monitoring to ensure staff continue to follow procedures. Effectively preventing central venous catheter-related bloodstream infections can enhance care quality and move healthcare closer to achieving the goal of zero tolerance.

The effectiveness of valerian acupressure on the sleep of ICU patients: a randomized clinical trial.

BACKGROUND: Severely ill patients often experience problems with sleep. Either acupressure or valerian aromatherapy are reported as helpful in promoting sleep. OBJECTIVES: The purpose of this study was to explore the effectiveness of valerian acupressure on the sleep of patients in the intensive care unit (ICU). DESIGN: A randomized clinical trial. SETTING: A 42-bed adult intensive care unit. PARTICIPANTS: Forty-one subjects in the experimental group and 44 subjects in the control group. METHODS: The measurement included observation, and actigraphy measures during 10 pm-6 am, and the Stanford Sleepiness Scale (SSS) measures on the next morning. Experimental groups received valerian acupressure on the Shenmen, Neiguan, and Yongquan acupoints between 7 pm and 10 pm of the second day while control groups received regular treatment. Heart rate was measured for 5 min before and after valerian acupressure present for HR variability analysis to measure relaxation response. RESULTS: The results indicated that after receiving valerian acupressure, patients' sleeping hours increased, wake frequency reduced and SSS grades declined. The HR variability data indicated relaxation response immediately after valerian acupressure. CONCLUSION: This study supports the hypothesis that valerian acupressure on the Shenmen, Neiguan, and Yongquan acupoints could improve the sleeping time and quality of ICU patients.

Pattern of expression and substrate specificity of chloroplast ferredoxins from Chlamydomonas reinhardtii.

Ferredoxin (Fd) is the major iron-containing protein in photosynthetic organisms and is central to reductive metabolism in the chloroplast. The Chlamydomonas reinhardtii genome encodes six plant type [Fe2S2] ferredoxins, products of PETF, FDX2-FDX6. We performed the functional analysis of these ferredoxins by localizing Fd, Fdx2, Fdx3, and Fdx6 to the chloroplast by using isoform-specific antibodies and monitoring the pattern of gene expression by iron and copper nutrition, nitrogen source, and hydrogen peroxide stress. In addition, we also measured the midpoint redox potentials of Fd and Fdx2 and determined the kinetic parameters of their reactions with several ferredoxin-interacting proteins, namely nitrite reductase, Fd:NADP+ oxidoreductase, and Fd:thioredoxin reductase. We found that each of the FDX genes is differently regulated in response to changes in nutrient supply. Moreover, we show that Fdx2 (Em = -321 mV), whose expression is regulated by nitrate, is a more efficient electron donor to nitrite reductase relative to Fd. Overall, the results suggest that each ferredoxin isoform has substrate specificity and that the presence of multiple ferredoxin isoforms allows for the allocation of reducing power to specific metabolic pathways in the chloroplast under various growth conditions.

FER1 and FER2 encoding two ferritin complexes in Chlamydomonas reinhardtii chloroplasts are regulated by iron.

Two unlinked genes FER1 and FER2 encoding ferritin subunits were identified in the Chlamydomonas genome. An improved FER2 gene model, built on the basis of manual sequencing and incorporation of unplaced reads, indicated 49% identity between the ferritin subunits. Both FER1 and FER2 transcripts are increased in abundance as iron nutrition is decreased but the pattern for each gene is distinct. Using subunit-specific antibodies, we monitored expression at the protein level. In response to low iron, ferritin1 subunits and the ferritin1 complex are increased in parallel to the increase in FER1 mRNA. Nevertheless, the iron content of the ferritin1 complex is decreased. This suggests that increased expression results in increased capacity for iron binding in the chloroplast of iron-limited cells, which supports a role for ferritin1 as an iron buffer. On the other hand, ferritin2 abundance is decreased in iron-deprived cells, indicative of the operation of iron-nutrition-responsive regulation at the translational or post-translational level for FER2. Both ferritin subunits are plastid localized but ferritin1 is quantitatively recovered in soluble extracts of cells while ferritin2 is found in the particulate fraction. Partial purification of the ferritin1 complex indicates that the two ferritins are associated in distinct complexes and do not coassemble. The ratio of ferritin1 to ferritin2 is 70:1 in iron-replete cells, suggestive of a more dominant role of ferritin1 in iron homeostasis. The Volvox genome contains orthologs of each FER gene, indicating that the duplication of FER genes and potential diversification of function occurred prior to the divergence of species in the Volvocales.

Somatic mutations at the trinucleotide repeats of androgen receptor gene in male hepatocellular carcinoma.

Androgen and androgen receptor (AR) have long been implicated in liver carcinogenesis, especially for the male dominance feature. However, whether AR gene could occur in somatic mutations that might contribute to this process has not yet been studied. DNA sequencing and genotyping were conducted for detecting the genetic aberrations of AR gene in 257 primary hepatocellular carcinomas (HCCs) and also the dysplastic nodules (DN) from another 11 patients. Twenty-one AR somatic mutations causing amino acid changes were identified in HCC and even in the precancerous DN. The missense somatic mutations of AR were rare in HCC (2 cases) but the trinucleotide repeat (TNR) changes, both at (CAG)n and (GGC)n, was a more common one (19 cases). Notably, all these mutations occurred in male patients and most TNR changes belonged to the contraction type (15 out of 19 cases, 78.9%), which has been reported to associate with increased AR transcriptional activity. Most samples with TNR changes did not show microsatellite instability, suggesting a different cause for these TNR mutations. Although no significant correlation was identified between AR mutations and the clinicopathologic parameters, we found the (CAG)n length significantly shorter in hepatitis B virus (HBV)(+) HCCs than in HBV(-) HCCs and the (GGC)n length significantly correlates with the overall survival. In conclusion, the mis-sense somatic mutations of AR were rare in HCC but the TNR change was a more common one, which exclusively occurred in males. Moreover, the length of TNR carried clinical significance in special HCC group.

[Applying self-regulation theory to develop an exercise protocol for diabetes mellitus].

The prevalence and mortality rates for diabetes in Taiwan have gradually increased in recent years. Moreover, according to the Bureau of National Health Insurance in 1998, diabetes accounts for 11.5% of overall health care spending in Taiwan, which imposes a substantial cost burden on society and patients. From the perspective of economics, exercise is the most effective, and yet the hardest behavior for patients to carry out in terms of diabetes control. Self-regulation theory is a model that can be used to develop effective strategies for behavioral modification. We designed activities for a diabetes exercise protocol on the basis of three components (i.e., self-monitoring, self-judgment and self-reaction) of self-regulation theory. It is expected that this protocol will effectively improve patients' exercise behavior.

Quantification and genotyping of hepatitis B virus in a single reaction by real-time PCR and melting curve analysis.

BACKGROUND/AIMS: Both viral titer and genotype of hepatitis B virus (HBV) play critical roles in determining clinical outcome and response to antiviral treatment in hepatitis B patients. In this study, a method was developed to determine both parameters in a single-tube reaction. METHODS: The method contains two consecutive steps, the first step used real-time PCR for quantification and second step used melting curve analysis for genotyping. For accurate quantification, the PCR primers and hybridization probes were selected from highly conserved regions to ensure the equivalent amplification and hybridization of all genotypes of HBVs. Within the sensor probe there exists signature single nucleotide polymorphisms (SNPs), which could effectively differentiate different HBV genotypes by showing different melting temperatures. RESULTS: The quantification results showed great consistency with the commercial assays in linear range from 10(2) to 10(11) copies/ml. By comparison with the traditional restriction fragment length polymorphism (RFLP) methods, 99% of samples were accurately genotyped by current assay, and with a higher detection rate. In addition, this method can detect mixed HBV infections. CONCLUSIONS: Currently, this methodology can be applied to areas prevalent with HBV genotypes B and C, providing an efficient alternative for clinical diagnosis and large-scaled longitudinal studies.

Allelic loss of chromosome 4q21 approximately 23 associates with hepatitis B virus-related hepatocarcinogenesis and elevated alpha-fetoprotein.

Allelic loss of chromosome 4q is one of the most frequent genetic aberrations found in human hepatocellular carcinoma (HCC) and suggests the presence of putative tumor suppressor genes within this region. To precisely define the region containing these tumor suppressor genes for further positional cloning, we tried a detailed deletion mapping strategy in 149 HCCs by using 49 microsatellite markers covering 4q12 approximately 25. A common region with allelic loss has been identified based on the interstitial deletions occurring within it; this region is found between D4S1534 and D4S1572 (a 17.5-cM genetic interval). When we included all cases with limited aberration regions for comparison, 2 smaller regions were derived: 1 between D4S1534 and D4S2460 (3.52 cM) and 1 between D4S2433 and D4S1572 (8.44 cM). A few candidate genes were found to be down-regulated in HCCs, but without sequence mutations. In these HCCs, 4q alleleic loss was associated with hepatitis B virus infection status and the elevation of serum alpha-fetoprotein (>/=400 ng/mL). In conclusion, the current study not only mapped a common allelic loss region on chromosome 4q, but it also revealed that its loss may be involved in hepatitis B virus-related hepatocarcinogenesis and the elevation of serum alpha-fetoprotein.

Changes in glucocorticoid and mineralocorticoid receptors of liver and kidney cytosols after pathologic stress and its regulation in rats.

OBJECTIVE: As effectors, glucocorticoid and mineralocorticoid receptors play an important role in pathologic stress. This study was designed to observe the changes in glucocorticoid receptor of liver cytosols and mineralocorticoid receptor of kidney cytosols after pathologic stress in rats. DESIGN: Controlled laboratory study. SETTING: Medical university. SUBJECTS: Male Wistar rats (weight range, 180-200 g). INTERVENTIONS: Rats received a low-degree or heavy-degree immersion scald that covered 10% or 35% total body surface area and were randomly divided to receive either tumor necrosis factor-alpha, interleukin-1beta polyclonal neutralizing antibody, alpha-melanocyte-stimulating hormone, KPV peptide (Ac-D-Lys-L-Pro-D-Val), or saline (control). The binding capacity and the apparent dissociation constant of the steroid-binding sites of normal, low-degree, and heavy-degree scalded rats were measured by radioligand-binding assay, with [3H]dexamethasone and aldosterone as the ligand, respectively. MEASUREMENTS AND MAIN RESULTS: The binding capacity of glucocorticoid receptor in hepatic cytosols in rats 12 hrs after heavy-degree scald (208.45 +/- 30.78 fmol/mg of protein) was lower than that of the control group (306.71 +/- 27.96 fmol/mg of protein; p < .01). The binding capacity of glucocorticoid receptor in hepatic cytosols in rats 12 hrs after low-degree scald (296.64 +/- 16.06 fmol/mg of protein) was not significantly different compared with the control group (p > .05). There were two types of mineralocorticoid receptor in kidney cytosols in rats, and their binding capacity and apparent dissociation constant were not identical. The binding capacity of mineralocorticoid receptor in rats 12 hrs after heavy-degree scald (binding capacity 1, 22.40 +/- 5.40 fmol/mg of protein; binding capacity 2, 196.30 +/- 32.50 fmol/mg of protein) was lower than that of the control group (binding capacity 1, 41.60 +/- 7.20 fmol/mg of protein; binding capacity 2, 317.60 +/- 70.00 fmol/mg of protein; p < .01). The binding capacity of mineralocorticoid receptor in kidney cytosols in rats 12 hrs after low-degree scald (binding capacity 1, 41.40 +/- 5.00 fmol/mg of protein; binding capacity 2, 314.80 +/- 45.70 fmol/mg of protein) was not significantly different compared with the control group (p > .05). The injections of anti-rat tumor necrosis factor-alpha, interleukin-1beta polyclonal neutralizing antibody, alpha-melanocyte-stimulating hormone, and KPV peptide (Ac-D-Lys-L-Pro-D-Val) might prevent a reduction in the binding capacity of glucocorticoid receptor in hepatic cytosols and mineralocorticoid receptor in kidney cytosols in rats with heavy-degree scald in vivo. CONCLUSIONS: These studies suggest that the glucocorticoid receptor of hepatic cytosols and the mineralocorticoid receptor of renal cytosols decreased in rats with heavy-degree immersion scald and that the injections of anti-rat tumor necrosis factor-alpha, interleukin-1beta polyclonal neutralizing antibody, alpha-melanocyte-stimulating hormone, and KPV peptide might increase the level of glucocorticoid receptor and mineralocorticoid receptor in vivo.