Forensic application of a novel MPS-based panel (90 STRs and 100 SNPs) in a non-exclusion parentage case with three autosomal STRs incompatibilities.

In kinship tests, the investigating of the forensic STRs usually provides decisive information to resolve relationship cases. We describe a parentage case with 3 genetic incompatibilities (D6S1043, D18S51 and D2S1338) between the child and alleged parent. With 90 STR loci and 100 SNP loci, the massively parallel sequencing (MPS)-based genotyping results support the certainty of parentage, and the mismatched alleles were considered to be mutations. MPS can provide additional allele sequence structures that can be used to infer the origins of the mutations. SNPs as supplementary markers can provide effective information to give an unequivocal statement of the parentage.

Metabolic acidosis in critically ill patients with cirrhosis: Epidemiology and short-term mortality risk factors.

BACKGROUND/AIMS: Metabolic acidosis is a common complication in patients with cirrhosis at the intensive care units (ICUs) and associated with increased mortality. The aim of our research was to explore the epidemiology and risk factors of metabolic acidosis in critically ill patients with cirrhosis. MATERIALS AND METHODS: A total of 975 patients with cirrhosis were selected into our study, and all participants were followed up for at least 28 days. Cox regression model and machine-learning algorithm were used to identify the importance of different risk factors, respectively. Finally, an improved prognostic model as Model for End-stage Liver Disease and metabolic acidosis (MELD-MA) was developed. RESULTS: Among the 975 patients with liver cirrhosis, 506 had metabolic acidosis, including 257 patients who had decompensated metabolic acidosis at ICU admission. The 28-day mortality was 41% (206/506) in patients with metabolic acidosis. Bilirubin (hazard ratio (HR): 1.023, 95% confidence interval (CI): 1.011-1.036), international normalized ratio (HR: 1.527, 95% CI: 1.332-1.750), pH (HR: 0.173, 95% CI: 0.047-0.640), BE-Lac (HR: 0.907, 95% CI: 0.868-0.948), and BE-Na (HR: 0.923, 95% CI: 0.859-0.991) were considered as independent prognostic parameters for 28-day mortality. MELD-NA had significantly higher discrimination (area under the receiver operating characteristic curve 0.79) than MELD and Child-Pugh score. CONCLUSION: Critically ill patients with cirrhosis have a high mortality rate and poor prognosis because of the high prevalence of metabolic acidosis. Lactic acidosis is the worst prognosis of all types of metabolic acidosis. MELD-MA performs well on the short-term mortality assessment in critically ill patients with cirrhosis and metabolic acidosis.

FMNL1 down-regulation suppresses bone metastasis through reducing TGF-ß1 expression in non-small cell lung cancer (NSCLC).

Approximately 40% of patients with non-small cell lung cancer (NSCLC) develop bone metastasis. The formin protein formin-like 1 (FMNL1) plays a key role in the pathogenic processes of hematopoietic malignancies, and has been reported to be associated with the progression of multiple types of cancer. In the study, we found that FMNL1 expression was markedly up-regulated in primary NSCLC samples, and stronger expression of FNML1 was detected in bone metastasis. Reducing FMNL1 expression significantly suppressed cell proliferation in NSCLC cells. We also investigated the functional effects of FMNL1 knockdown on the inhibition of migration and invasion by meditating the expression of epithelial to mesenchymal transition (EMT)-associated signals in NSCLC cells. The transforming growth factor-ß1 (TGF-ß1)/SMADs signaling pathway was repressed in FMNL1-knockdown NSCLC cells. Further studies indicated that additional treatment with TGF-ß1 could markedly abrogate FMNL1 knockdown-induced suppression of migration and invasion in NSCLC cells. In addition, NSCLC cell-induced osteoclastogenesis was also inhibited by FMNL1 deletion, as evidenced by the down-regulated expression of tartrate-resistant acid phosphatase (TRAP) and NFATc1. In vivo studies confirmed the results that FMNL1 knockdown markedly limited tumor growth. Importantly, decreasing FMNL1 reduced bone metastasis ability in vivo. Therefore, our results demonstrated that suppressing FMNL1 expression could inhibit bone metastasis in NSCLC through blocking TGF-ß1 signaling, and FMNL1 might be a novel target for developing effective therapeutic strategy to limit the bone metastasis of NSCLC.

Characterization of polyethylene glycol-polyethyleneimine as a vector for alpha-synuclein siRNA delivery to PC12 cells for Parkinson's disease.

AIMS: Gene therapy targeting the SNCA gene yields promising results in the treatment of Parkinson's disease (PD). The most challenging issue of the RNAi gene therapy strategy is maintaining efficient delivery without inducing significant toxicity and other adverse effects. This study aimed to characterize polyethylene glycol-polyethyleneimine as a vector for alpha-synuclein siRNA delivery to PC12 cells for Parkinson's disease. METHODS: The characteristics of PEG-PEI/siSNCA were analyzed via gel retardation assay and assessments of particle size and zeta potential. MTT cytotoxicity assay and flow cytometry were used to detect cytotoxicity and transfection efficiency in PC12 cells. Confocal laser scanning microscopy was employed to examine the intracellular distribution of PEG-PEI/FITC-siSNCA after cellular uptake. RT-PCR and western blotting were used to measure SNCA expression. The MTT cytotoxicity assay was used to study the effect of PEG-PEI/siSNCA on cell viability. The protective effect of PEG-PEI/siSNCA on MPP+-induced apoptosis in PC12 cells was examined via flow cytometry and Hoechst staining. RESULTS: PEG-PEI/siSNCA complexes were well-developed; they exhibited appropriate particle sizes and zeta potentials at a mass ratio of 5:1. In vitro, PEG-PEI/siSNCA was associated with low cytotoxicity and high transfection efficiency. Complexes were capable of successfully delivering siSNCA into PC12 cells and releasing it from the endosome. Furthermore, PEG-PEI/siSNCA could effectively suppress SNCA mRNA expression and protected cells from death via apoptosis induced by MPP(+) . CONCLUSIONS: Our results demonstrate that PEG-PEI performs well as a vector for alpha-synuclein siRNA delivery into PC12 cells. Additionally, PEG-PEI/siSNCA complexes were suggested to be able to protect cells from death via apoptosis induced by MPP(+) . These findings suggest that PEG-PEI/siSNCA nanoparticles exhibit remarkable potential as a gene delivery system for Parkinson's disease.

[Nosocomial infection caused by Pseudomonas aeruginosa in intensive care unit].

OBJECTIVE: To investigate the risk factors of nosocomial infection caused by Pseudomonas aeruginosa in intensive care unit (ICU), in order to provide reference for an effective measure of infection control. METHODS: A retrospective study of cases of Pseudomonas aeruginosa infection occurring in ICU was made with multivariable Logistic regression analysis. The clinical data of 1 950 cases admitted from January 2002 to December 2006 were found to have nosocomial infection caused by Pseudomonas aeruginosa were analyzed in order to identify its independent risk factors. RESULTS: Sixty-four out of 1 950 patients were found to suffer from nosocomial infection caused by Pseudomonas aeruginosa, the morbidity rate was 3.3%. At the same time, and in the same department, 37 patients suffering from infection caused by Escherichia coli, served as control group. Univariate analysis showed that the risk factors for nosocomial infection caused by Pseudomonas aeruginosa were the use of corticosteroid, unconsciousness or craniocerebral trauma, abdominal surgery, thorax/abdomen drainage tube, mechanical ventilation, and tracheostomy [the use of corticosteroid: odds ratio (OR)=3.364, 95% confidence interval (95%CI) 1.445-7.830; unconsciousness or craniocerebral trauma: OR=4.026, 95%CI 1.545-10.490; abdominal surgery: OR=0.166, 95%CI 0.068-0.403; thorax/abdomen drainage tube: OR=0.350, 95%CI 0.150-0.818; tracheostomy: OR=4.095, 95%CI 1.638-10.740]. Multivariate analysis showed that the independent risk factors of nosocomial infection caused by Pseudomonas aeruginosa in ICU were: the use of corticosteroid and mechanical ventilation [the use of corticosteroid: OR=3.143, 95%CI 1.115-8.856; mechanical ventilation: OR=3.195, 95%CI 1.607-6.353, P<0.05 and P<0.01]. CONCLUSION: The independent risk factors of nosocomial infection caused by Pseudomonas aeruginosa in ICU are the use of corticosteroid and mechanical ventilation. Measures should be taken to take care of the risk factors in order to prevent nosocomial infection caused by Pseudomonas aeruginosa in ICU.

Influence of memory effect caused by aged bag filters on the stack PCDD/F emissions.

To our best knowledge, this study is the first research which investigates whether the PCDD/F memory effect could also be caused by bag filters. In this study, the PCDD/F characteristics of the flue gases in front of and behind the bag filters of one municipal solid waste incinerator (MSWI) and two electric arc furnaces (EAFs) were investigated to clarify the memory effect of PCDD/Fs and their influence on emissions. After the bag filters were operated for over four years, the PCDD/F concentrations in the stack flue gases were all higher than those prior to the aged bag filter, rendering a negative PCDD/F removal. This memory effect is because gaseous phase PCDD/Fs are released from the contaminated filters of the aged bag filters. The release of 2,3,7,8-TeCDF, 1,2,3,7,8-PeCDF and 2,3,4,7,8-PeCDF from the contaminated filters of the EAFs increased the PCDD/F concentration and their fractions in the stack flue gases. In contrast, the MSWI exhibited increasing fractions of 1,2,3,4,6,7,8-HpCDD, OCDD after flue gases passed through its aged bag filter. The activated carbon of the MSWI adsorbed more lower chlorinated-substituted PCDD/Fs in the raw flue gases, and the remaining higher chlorinated congeners gradually accumulated on the aged filters, and were released in lower PCDD/F concentration conditions.

A new inflammation marker of chronic obstructive pulmonary disease-adiponectin.

BACKGROUND: This study was undertaken to measure the concentration of adiponectin (APN) in serum and induced sputum in patients with chronic obstructive pulmonary disease (COPD during acute exacerbation (AECOPD) and at stable stage and to determine the role of APN as a marker of inflammation in the pathogenesis of COPD. METHODS: All the patients in this prospective study were enrolled from October 2008 to October 2009, including 30 male AECOPD patients from the emergency department, 30 male stable COPD patients from the department of respiratory diseases, and 30 healthy non-smoking male controls from the department of medical examination. The serum and induced sputum were collected from each patient. All of the patients had normal weight (BMI range 18.5-24.9 kg/m(2)). Patients with severe bronchial asthma, bronchiectasis or autoimmune disease were excluded. Cell count and classification was performed for the induced sputum. The concentrations of APN, IL-8, IL-6 and TNF-α were measured by ELISA. Pulmonary function was tested among the three groups. Comparisons between the groups were conducted by Student's t test, ANOVA analysis or nonparametric test. Correlation analysis was carried out by Pearson's product-moment correlation coefficient test or Spearman's rank-order correlation coefficient test. RESULTS: The concentrations of APN in the serum or induced sputum in AECOPD patients were significantly higher than those in stable COPD patients or healthy non-smoking controls (P<0.01). The concentration of APN in stable COPD patients was significantly higher than that in healthy non-smoking controls (P<0.01). For the AECOPD patients, APN was positively correlated with IL-8 and TNF-α in the serum and induced sputum (r=0.739, 0.734, 0.852, 0.857 respectively, P<0.05). For the stable COPD patients, APN was also positively correlated with IL-8 and TNF-α in the serum and induced sputum (r=0.751, 0.659, 0.707, 0.867 respectively, P<0.05). In addition, for the AECOPD patients, APN was positively correlated with the percentage of neutrophils in the induced sputum (r=0.439, P<0.05). CONCLUSIONS: APN is involved in the process of systematic and airway inflammation of COPD. This process is related to neutrophils in the airway, IL-8 and TNF-α. APN could be used as a new marker for inflammation of COPD.

[Effect of opening of neuronal mitochondrial permeability transition pore on respiratory function after cardiopulmonary resuscitation in rats].

OBJECTIVE: To investigate the effect of opening of neuronal mitochondrial permeability transition pore (MPTP) on respiratory function after cardiopulmonary resuscitation (CPR) in rats and its possible mechanism. METHODS: Cardiac arrest (CA)/CPR rat model was reproduced by asphyxiation and ice-cold KCl followed resuscitation and restoration of spontaneous circulation (ROSC). The rats were sacrificed by decapitation at 3, 6, 12, 24, 48 and 72 hours. Isolation of brain cortex neuronal mitochondria was processed. MPTP opening degree was examined by spectrophotometer. Clark oxygen electrode was used to measure mitochondrial respiratory function: the mitochondrial ultra structure was examined with transmission electron microscope (TEM). RESULTS: Mitochondrial respiratory function was severely injured after CA/CPR. Mitochondrial respiratory state III (R3) was decreased. Neural cell MPTP opened persistently after ROSC. The opening degree of MPTP did not reach the peak instantly, and its change depended on time. It remained at a low level within 6 hours after ROSC, then rapidly opened, reaching the maximal degree at 12 hours, but it became smaller at 24 hours. At 48 hours the degree of opening became larger again, but shrank once more at 72 hours. However, it did not reach the normal level (all P<0.05). Although R3 was decreased, mitochondrial respiratory state IV (R4) was increased, meanwhile the respiratory control rate (RCR) and P/O ratio descended markedly. They maintained at low levels along with the elapse of time (P<0.05 or P<0.01). TEM revealed obvious injury to neurons. Correlation analysis showed that the MPTP opening degree and RCR was obviously positively correlated (r=0.025, P<0.05). CONCLUSION: The opening of MPTP is the main cause of aggravation of energy metabolism disturbance of neural cells after CPR. To take measures to inhibit the opening of MPTP within 12 hours after ROSC may promote improvement of neuronal mitochondrial function, and it might help win the chances for neural function to recover.

First aid strategy for severe traumatic patients in hospital.

OBJECTIVE: To study the emergency management principles of severe trauma in hospital (injury severity score larger than or equal to 16). METHODS: We used "ATP principle" to manage severe traumatic patients. The ATP principle is composed of: 1) attending surgeons offering initial management (A); 2) teamwork commencement immediately after patients admitted to hospital (T); 3) parallel principle, ie, emergency resuscitation, evaluation and laboratory test performed simultaneously (P). Clinical effects before and after applying ATP principle were retrospectively analyzed and compared. RESULTS: During January 1, 2002 to December 31, 2003, 338 patients were treated without applying ATP principle, in which ISS was 25.9+/-6.4, 152 cases died with the mortality being 39.2%, and the time stayed in emergency department and the time to operation room after admission were (102.8+/-16.7) min, (140.3+/-20.6) min, respectively. During January 1, 2004 to December 31, 2005, 438 patients were treated based on ATP principle, in which ISS was 28.6+/-7.8, 87 cases died with the mortality being 19.9%, and the time in emergency department and the time to operation room after admission were (69.5+/-11.5) min, (89.6+/-9.3) min, respectively. ISS showed no significant difference between the two groups (P larger than 0.05) but the mortality, the time stayed in emergency department and the time to operation room after admission were greatly reduced and showed significant difference between the two groups (P less than 0.05). CONCLUSIONS: Applying ATP principle to treat severe traumatic patients can shorten emergency treatment time in hospital and decrease mortality.

[Effects of rhubarb and different routes of nutrition support on invasive fungal infection].

OBJECTIVE: To study the effects of rhubarb and the different routes of nutrition support on invasive fungal infection. METHODS: One thousand and ninety patients, who suffered from sepsis subsequent to trauma, shock and infection were enrolled in this study. The patients were randomly divided into two groups, 637 cases in rhubarb preventive treatment group and 453 cases in non-preventive rhubarb treatment group. They were again divided into four subgroups: enteral nutrition support, and no nutrition support group. The incidence of invasive fungal infection was observed in those groups. RESULTS: The incidence of invasive fungal infection in rhubarb preventive treatment group (3.0%) was much lower than that in non-preventive rhubarb treatment group (11.5%). There was significant difference between two groups (P<0.05). Furthermore, fewer patients developed invasive fungal infection in enteral nutrition support and enteral combined parenteral nutrition support subgroups after preventive rhubarb treatment (0.9% and 2.1%), compared with parenteral nutrition support and no nutrition support subgroups (30.4% and 61.3%) and corresponding subgroups with non-preventive treatment of rhubarb (3.9% and 7.1%, P<0.05 or P<0.01). In addition, the route of nutrition support also affected the incidence of invasive fungal infection. Patients in enteral nutrition support and enteral combined parenteral nutrition support subgroups had lower incidence of invasive fungal infection than in parenteral nutrition support and no nutrition support subgroups (all P<0.05), and the incidence was the highest in no nutrition support subgroup. There were no significant difference between parenteral nutrition support and no nutrition support subgroups. CONCLUSION: Rhubarb and enteral nutrition support have preventive effects on invasive fungal infection via gut mechanism.

Influence of start-up on PCDD/F emission of incinerators.

This study aims to evaluate the influence of start-up on polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) concentration in the stack flue gas of incinerators and its contributing PCDD/F emission. The PCDD/F emission of the first sample among three consecutive stack flue gas samples of five intermittent incinerators, which sampled at a stable combustion condition after start-up, is 2-3 times higher than the mean of the others. For verifying the PCDD/F characteristics of incinerators during start-up, one continuous MSWI was investigated for two years. The elevated PCDD/F emissions of the MSWI during start-up could reach 96.9 ng I-TEQN m(-3) and still maintained a high PCDD/F emission (40 times higher than the Taiwan emission limit) even 18 h after the injection of activated carbon, indicating the memory effect. Taking the MSWI for example, which consists of four incinerators, the estimated annual PCDD/F emission from normal operational conditions was 0.112 g I-TEQ. However, one start-up procedure can generate approximately 60% of the PCDD/F emissions for one whole year of normal operations. And the PCDD/F emission, which is the result of the start-ups of four incinerators, was at least two times larger than that of a whole year's normal operations, without consideration for the PCDD/F emission contributed by the long lasting memory effect.

[Preventive effect and clinical significance of garlicin injection on mechanical ventilator-associated low respiratory tract deep-seated fungal infection].

OBJECTIVE: To investigate the preventive effect and clinical significance of garlicin injection on ventilator-associated low respiratory tract deep-seated fungal infection (VRFI) as pneumonia. METHODS: Retrospectively analysis on 147 patients underwent mechanical ventilation in our Intensive Care Unit (ICU) in recently 3 years was conducted. According to the garlicin injection administration was used or not, 79 patients with WVUH (West Virgina University Hospital) score > or = 25 points were selected and divided into the preventive group and the control group, and the differences in baseline conditions, incidence of VRFI and hospital outcome between the two groups were compared. RESULTS: There was no significant difference between the two groups in baseline data as age, sex, APACHE II score, SAPS II score and WVUH score. But the incidence of VRFI in the preventive group was significantly lower than that in the control group (P<0.01). The relative risk in the latter was 2.06 (95% CI, 1.25-3.38). No significant difference was found between the two groups in hospital mortality (P>0.05). CONCLUSION: Garlicin injection can effectively reduce the incidence of VRFI, and is inexpensive, it is an ideal prophylactic anti-fungal drug.

[Diagnostic effect of bronchoalveolar lavage in early lung injury caused by enterogenic infection in rat].

OBJECTIVE: To study the diagnostic effect of bronchoalveolar lavage in early lung injury by observing changes in inflammatory mediators in early lung injury caused by enterogenic infection. METHODS: Eighty-four Sprague-Dawley rats were randomly divided into infection group and sham-operation group. Cecal ligation and perforation was utilized to produce abdominal infection in rats. Six groups were sacrificed respectively at 0, 24, 48, 72, 96, 120 hours after operation. The differential cell count in bronchoalveolar lavage fluid (BALF) was assessed. The concentrations of endotoxin, phospholipase A2 (PLA2) and tumor necrosis factor-alpha (TNF-alpha) in BALF, lung and plasma were assayed. RESULTS: The neutrophil percentage of BALF increased progressively. The concentrations of endotoxin, PLA2 and TNF-alpha in BALF, lung and plasma were significantly increased. The levels of endotoxin and PLA2 in lung tissue were respectively correlated positively with those in BALF and plasma (BALF and lung: r=0.904, P<0.05; BALF and plasma: r=0.895, P<0.05; lung and plasma: r=0.946, P<0.01). Significant positive correlation was also present between the TNF-alpha levels in BALF and lung (r=0.952 P<0.01), but not between the TNF-alpha level in plasma and that in lung or BALF (r=0.684, r=0.608, both P>0.05). CONCLUSION: The examinations of bronchoalveolar lavage may help discover early lung injury caused by enterogenic infection.