Genetic variant rs72613567 of HSD17B13 gene reduces alcohol-related liver disease risk in Chinese Han population.

BACKGROUND & AIMS: Recently, the variant rs72613567:TA in the 17-beta-hydroxysteroid dehydrogenase 13 (HSD17B13) has been associated with reduced levels of ALT and AST and a reduced risk of alcohol-related liver disease (ALD) in the European population. Therefore, the aim of this study was to investigate the association between the polymorphisms of HSD17B13 and ALD, liver serum markers and patatin-like phospholipase domain-containing protein 3 (PNPLA3) p.I148M in the Chinese Han population. METHODS: A case-control study was performed from five centres and included 769 ALD patients and 767 healthy controls. Two SNPs (rs72613567 and rs6834314) in HSD17B13 were genotyped using the Sequenom MassArray system and allele association analysis was performed using PLINK 1.90 software. RESULTS: HSD17B13 rs72613567:TA allele was associated with a reduced risk of ALD by 19% (95% confidence interval [CI]: 0.05-0.31, P = .01), uniformly, the G allele in the rs6834314 reduced the risk of ALD by 19% (95% CI: 0.05-0.31, P = 8.28 × 10-3). And the genotypes of two SNPs were associated with reducing the risk of ALD in three genetic model analysis. In addition, we found that TA allele was associated with lower levels of serum ALT, AST and GGT (P = .005, .007 and .02, respectively), higher level of serum ALB (P = .02), but not associated with ALP. In this cohort, the interaction between HSD17B13 rs72613567 and the steatogenic allele PNPLA3 rs738409 was not validated. CONCLUSION: The present study revealed that HSD17B13 rs72613567 was significantly associated with a reduced risk of ALD in Chinese Han population.

Application Value of Mass Spectrometry in the Differentiation of Benign and Malignant Liver Tumors.

BACKGROUND Differentiation of malignant from benign liver tumors remains a challenging problem. In recent years, mass spectrometry (MS) technique has emerged as a promising strategy to diagnose a wide range of malignant tumors. The purpose of this study was to establish classification models to distinguish benign and malignant liver tumors and identify the liver cancer-specific peptides by mass spectrometry. MATERIAL AND METHODS In our study, serum samples from 43 patients with malignant liver tumors and 52 patients with benign liver tumors were treated with weak cation-exchange chromatography Magnetic Beads (MB-WCX) kits and analyzed by the Matrix-Assisted Laser Desorption Time of Flight Mass Spectrometry (MALDI-TOF-MS). Then we established genetic algorithm (GA), supervised neural networks (SNN), and quick classifier (QC) models to distinguish malignant from benign liver tumors. To confirm the clinical applicability of the established models, the blinded validation test was performed in 50 clinical serum samples. Discriminatory peaks associated with malignant liver tumors were subsequently identified by a qTOF Synapt G2-S system. RESULTS A total of 27 discriminant peaks (p<0.05) in mass spectra of serum samples were found by ClinPro Tools software. Recognition capabilities of the established models were 100% (GA), 89.38% (SNN), and 80.84% (QC); cross-validation rates were 81.67% (GA), 81.11% (SNN), and 86.11% (QC). The accuracy rates of the blinded validation test were 78% (GA), 84% (SNN), and 84% (QC). From the 27 discriminatory peptide peaks analyzed, 3 peaks of m/z 2860.34, 2881.54, and 3155.67 were identified as a fragment of fibrinogen alpha chain, fibrinogen beta chain, and inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), respectively. CONCLUSIONS Our results demonstrated that MS technique can be helpful in differentiation of benign and malignant liver tumors. Fibrinogen and ITIH4 might be used as biomarkers for the diagnosis of malignant liver tumors.

Clinical Features of Patients With Ebola Virus Disease in Sierra Leone.

BACKGROUND: Clinical and laboratory data available on patients with Ebola virus disease (EVD) remain extremely limited. We summarized the clinical characteristics of patients with EVD and analyzed the factors related to their death. METHODS: Patients admitted for care at the Freetown China-Sierra Leone Friendship Hospital during 1 October-14 November 2014 were enrolled in this study. The clinical data of these patients were retrospectively analyzed. RESULTS: Sixty-one patients were confirmed to have EVD; 28 of them (45.9%) were male and 33 (54.1%) were female. Their median age was 28 years (range, 1.17-67 years). The median duration from symptom onset to clinic visit time was 5 days (range, 1-16 days). Among these patients, 42 of them (68.9%) died. Of the confirmed cases, 18.0% did not present with fever. Patients aged >30 years had a higher fatality rate than those <30 years (87.0% vs 57.9%; P = .0175). The mean duration from symptom onset to clinic presentation of the survivors (4.57 ± 2.79 days) was shorter than that of the nonsurvivors (6.34 ± 3.33 days). Clinical factors associated with a fatal outcome included weakness, extreme fatigue, vomiting, diarrhea, mental symptoms, bleeding, and loss of appetite. No statistical difference in the case fatality rate between males and females was found (P = .2061). CONCLUSIONS: The mortality of patients with EVD was closely associated with age and duration from symptom onset to presentation for care. Patients with EVD identified in the current outbreak did not necessarily have fever. Early diagnosis of the disease and timely symptomatic treatment may greatly contribute to the reduction of fatality rate of patients with EVD.

Laboratory features throughout the disease course of influenza A (H1N1) virus infection.

BACKGROUND: Influenza has emerged every year but a complete profile of laboratory indices throughout the disease course remains unknown. METHODS: Clinical data was collected from 28 confirmed cases of the pandemic influenza H1N1 2009. The levels of serum iron (Fe), carbon dioxide combining power (CO2-CP), total complement hemolytic activity (CH50), C-reactive protein (CRP), and white blood cell (WBC) and differential count were analyzed. RESULTS: Major laboratory abnormalities recokled for patients upon admission were lymphopenia (96.4%), eosinopenia (50.0%), hypoferremia (92.9%), decreased levels of serum CO2-CP (60.7%), increased levels of serum CRP (84.6%) and serum CH50 (71.4%). The serum iron and CO2-CP concentration and the counts for lymphocytes, eosinophils, and basophils were significantly increased four days after sickness was noticed compared with the first three days of illness (p < 0.05). The total WBC and neutrophil counts were significantly decreased four days after onset of illness compared with the counts over the first three days (p < 0.05). The monocyte count and CRP concentration was significantly decreased 7 days after onset of illness compared with first 3 days after illness onset (p < 0.05). The serum CH50 concentrations were higher than the normal range during disease course and significantly elevated 7 days after onset of illness compared with the first 6 days after illness onset (p < 0.05). CONCLUSIONS: The serum levels of iron, CO2-CP, CH50, CRP, and WBC and differential count Were significantly varied during the whole pandemic influenza (H1N1) 2009. The development of WBC count in patients with influenza may be an effective predictor for severity of illness.

[A clinical and pathological analysis of 22 cases of primary sclerosing cholangitis].

OBJECTIVE: To characterize the clinical, laboratory, imaging and pathological features of primary sclerosing cholangitis (PSC) and investigate the impact of ursodeoxycholic acid (UDCA) therapy on patient prognosis. METHODS: The medical records of 22 patients diagnosed with PSC between 2002 and 2011 were retrospectively reviewed. The PSC diagnosis had been made in patients with suspect biochemical abnormalities following evaluation by magnetic resonance cholangiopancreatography (MRCP) and/or endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC). Fibrosis and inflammation were assessed by immunohistochemical analyses of tissue biopsies. Outcome of patients treated with UDCA (13-15 mg/kg/day, oral) were compared to that of patients without UDCA treatment by the X2 or corrected X2 tests. RESULTS: Among the 22 PSC patients, the majority was male (n=15) and presented with fatigue, dark urine, and body weight loss (n=15). Four cases had ulcerative colitis. At admission, all 22 cases showed elevated levels of alkaline phosphatase[ALP: (348+/-184) U/L], 19 cases showed elevated alanine aminotransferase [ALT: (94.0+/-67.0) U/L] and aspartate aminotransferase [AST: (98.0+/-67.0) U/L], and 15 cases showed elevated levels of total bilirubin (99.0+/-115.0) mumol/L and direct bilirubin (74.4+/-92.4 mumol/L. ERCP examination showed segmental intrahepatic bile duct stenosis with expansion, and stiff and enlarged gallbladder bile ducts, but unclear findings for the common bile ducts and pancreatic ducts. MRCP showed beading of the intrahepatic bile duct, stiffness of the bile duct wall, and dilation of the common bile duct. Fibrosis and inflammation were observed in the bile ducts, along with hyperplasia and the typical features of "onion skin" fibrosis and fibrous obliterative cholangitis. Five of the 10 patients treated with UDCA improved, and seven of the 12 patients in the non-UDCA treatment group improved. There was no statistically significant difference in outcome between the groups (paired X2=0.333, corrected X2=0.083, P more than 0.05). CONCLUSION: PSC patients were predominantly male and the common clinical manifestations were fatigue, dark urine, and body weight loss. At admission, serum biochemical indicators of cholangitis were increased significantly and subsequent imaging studies confirmed the suspected diagnosis by showing obvious characteristic changes. UDCA treatment did not significantly improve patient prognosis.

Immunologic changes during Pandemic (H1N1) 2009, China.

We analyzed changes in immunologic values over time for 28 hospitalized patients with pandemic (H1N1) 2009. Levels of interleukin-6, interferon-y, and interleukin-10 increased 1 day after illness onset and then decreased to baseline levels. Levels of virus-specific antibody were undetectable 1 day after illness onset and peaked 36 days later.

Nasal colonization of and clonal transmission of methicillin-susceptible Staphylococcus aureus among Chinese military volunteers.

Military facilities provide unique opportunities for studying Staphylococcus aureus nasal colonization and transmission patterns. In this cross-sectional observational study, we assessed the prevalence of S. aureus nasal colonization among Chinese military volunteers in two camps in the Beijing area. Antimicrobial resistance patterns, risk factors for colonization, and transmission patterns using pulsed-field gel electrophoresis were also evaluated. From May to July 2007, 1,044 nasal swabs were collected from military volunteers from suburban (560) and urban (484) camps. A total of 209 S. aureus isolates were recovered, of which all were methicillin susceptible. Independent factors associated with methicillin-susceptible S. aureus (MSSA) nasal colonization included younger age (odds ratio [OR] = 1.51, 95% confidence interval [95% CI] = 1.03 to 2.21, P = 0.0347), higher education (OR = 1.38, 95% CI = 1.10 to 1.73, P = 0.0056), shorter length of service (OR = 1.74, 95% CI = 1.28 to 2.36, P = 0.0004), nonsmoking (OR = 1.61, 95% CI = 1.14 to 2.28, P = 0.0069), and inactive participation in social events (OR = 2.40, 95% CI = 1.25 to 5.49, P = 0.0082). Among 209 MSSA isolates, 126 (60.3%) were determined to be epidemic and a total of 12 genotypes were identified, of which four (90 isolates [71.4%]) represented the majority of strains. Length of service and camp location were statistically related to the four major MSSA genotype clonal transmissions. Our data indicated that MSSA, not methicillin-resistant S. aureus (MRSA), nasal colonization and clonal transmission occur in healthy military volunteers in Beijing. Younger, female, nonsmoking volunteers with higher education, little or no participation in social events, and less time in service are at higher risk for nasal MSSA carriage.

[The distribution and antimicrobial resistance tendency of pathogens associated with diarrhea in Beijing].

OBJECTIVE: To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study, vaccination research and clinical treatment. METHODS: Enteric pathogenic bacteria were cultured and identified to species, group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. RESULTS: Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp. (75.11%) was the most frequently isolated pathogens and followed by Vibrio spp. (12.7%), Salmonella spp. (6.28%), Aeromonas spp. (4.43%) and Escherichia coli (1.25%). During the period from 1994 to 2005, diarrheal pathogens had a trend of decrease especially Shigella spp. and Salmonella spp. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S. sonnei, S. dysenteriae and S. boydii constituted 23.98%, 0.22% and 0.01% respectively. The sensitivity of different species, group or serotype to different antimicrobial agents was not the same. S. flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S. sonnei and Vibrio spp. had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. CONCLUSION: There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes, so strict surveillance is always needed.

Single-nucleotide rs738409 polymorphisms in the PNPLA3 gene are strongly associated with alcoholic liver disease in Han Chinese males.

OBJECTIVE: Alcoholic liver disease (ALD) is a chronic liver disorder caused by the consumption of large amounts of alcohol. Genome-wide association studies have recently confirmed that polymorphisms in PNPLA3 predispose individuals to ALD and have identified risk loci of MBOAT7 and TM6SF2 in persons of European descent. However, the association with alcoholic liver damage has not been evaluated thus far in a Han Chinese population. METHODS: We performed a large case-control multicenter study of 507 ALD patients and 645 ethnically matched healthy controls. Five SNPs were genotyped using matrix-assisted laser desorption/ionization time of flight mass spectrometry, and association analysis was performed using PLINK 1.07 software. RESULTS: The rs738409 in the PNPLA3 gene was found to be significantly associated with ALD in allele and genotype frequencies (p = 6.25 × 10-14 and p = 9.05 × 10-13). The frequencies of the risk allele G in rs738409 were notably higher in ALD compared to controls (odds ratio = 1.93, 95% confidence interval = 1.63-2.28). The current study showed that the genotype frequencies of three genetic models were also statistically significant (p = 1.07 × 10-13, p = 9.3 × 10-8, and p = 1.57 × 10-12). Additionally, the G-allele of rs738409 was associated with a variety of clinical manifestations such as elevated alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), and mean corpuscular volume (MCV) in the patients with ALD. CONCLUSION: In a Han Chinese population, the present study confirmed that PNPLA3 polymorphism rs738409 was more likely to influence the susceptibility to ALD. However, no statistically significant differences for the allele and genotype frequencies of rs626283, rs641738 in MBOAT7, rs10401969 in SUGP1 and rs58542926 in TM6SF2 were found between ALD patients and healthy controls.

ITIH4: Effective Serum Marker, Early Warning and Diagnosis, Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is a highly lethal malignant tumor evolved from cirrhosis. It is quite significant to seek accurate, easy markers for early warning and diagnosis of HCC. Through prospective cohort follow-up study and mass spectrometry, we discovered and verified a serum marker valuable for early warning and diagnosis. Follow-up observation was performed on cirrhosis patients. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was adopted to detect the serums of patients, and the serum polypeptides with a potential value in early HCC warning and diagnosis were screened. Electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF-MS/MS) was exploited to identify these screened polypeptides. Moreover, the serum marker concentration was determined by ELISA to validate the clinical value of the serum marker. Among 109 cirrhosis patients followed up for two years, 29 patients (26.6%) finally progressed into HCC. MALDI-TOF MS shows that the concentration of a 3155.66Da polypeptide was significantly different between the patients that progressed into HCC and those not. Through MS/MS identification, it is confirmed that the polypeptide is inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4). The serum ITIH4 concentrations in two groups were measured with ELISA and compared with Alpha-fetoprotein (AFP). Results show that serum ITIH4 and AFP concentrations were negatively correlated (r=-0.263, p=0.0006), and the ITIH4 concentration had a significant intergroup difference (p=0.000). Receiver operating characteristic (ROC) curve indicates that its predictive value (area under the curve, AUC) is 0.667, superior to AFP. For the patients progressing into HCC, serum samples were separately collected when they were recruited and diagnosed as cirrhosis. Measurement on these samples reveals that ITIH4 was declining during the progression of HCC (p=0.006). By virtue of mass spectrometry, we discovered and identified a biomarker valuable for early HCC warning and diagnosis. This marker overperforms the commonly used AFP, demonstrating a bright prospect.

A rapid point-of-care test for dengue virus-1 based on a lateral flow assay with a near-infrared fluorescent dye.

Dengue fever is caused by the dengue virus (DENV), and DENV1 is the prevalent epidemic serotype in south China. A new lateral flow assay (LFA) based on a near-infrared (NIR) fluorescent dye was developed to detect anti-DENV1 IgG antibodies. DyLight-800 was used as the marker conjugated to goat anti-human IgG antibodies, and recombinant dengue type 1 envelope protein was used as the capture protein on the test line. Twenty samples from patients infected with DENV1 and 160 negative controls were analyzed using this new NIR-LFA. The results of the NIR-LFA were compared with the results of Panbio Dengue IgG ELISA and the Dengue Duo IgM/IgG Cassette. Nineteen confirmed DENV1-positive samples were identified by NIR-LFA, giving 95% (19/20) sensitivity. No significant differences existed in the results when the 20 primary clinical samples were analyzed using NIR-LFA, Panbio ELISA, and the Dengue Duo Cassette. However, NIR-LFA had a lower limit of detection than IgG ELISA and Duo IgM/IgG Cassette did when analyzing a 2-fold dilution series of the 19 samples positively identified by NIR-LFA. When incorporated with an NIR POCT device, the new NIR-LFA was rapid, easy to use, and highly sensitive in detecting DENV1, and has potential for application to clinical diagnosis.

Artificial neural network models for early diagnosis of hepatocellular carcinoma using serum levels of α-fetoprotein, α-fetoprotein-L3, des-γ-carboxy prothrombin, and Golgi protein 73.

More than 70% of hepatocellular carcinoma (HCC) cases develop as a consequence of liver cirrhosis (LC). Here we have evaluated the diagnostic potential of four serum biomarkers, and developed models for HCC diagnosis and differentiation from LC patients. Serum levels of α-fetoprotein (AFP), AFP-L3, des-γ-carboxy prothrombin (DCP), and Golgi protein 73 (GP73) were analyzed in 114 advanced HCC patients, 81 early stage HCC patients, and 152 LC patients. Multilayer perceptron (MLP) and radial basis function (RBF) neural networks were used to construct the diagnostic models. Using all stages, HCC diagnostic models had a higher sensitivity (>70%) than the individual serum biomarkers, whereas only early stage HCC diagnostic models had a higher specificity (>80%). The early stage HCC diagnostic models could not be used as HCC screening tools due to their low sensitivity (about 40%). These results suggest that a combination of the two models might be used as a screening tool to distinguish early stage HCC patients from LC patients, thus improving prevention and treatment of HCC.

The Clinical Values of Serum Markers in the Early Prediction of Hepatocellular Carcinoma.

The early prediction values of diagnostic markers for hepatocellular carcinoma (HCC) are still unclear at present. This study evaluated the prediction value of ten serum markers in HCC. A total of 109 cases of hepatic cirrhosis patients were followed up for 36 months and the relationship between the lifetime risk of developing HCC and levels of serum markers was analyzed. 31.2 (34/109) percent of hepatic cirrhosis patients developed HCC during the study's timeframe. Higher alpha-fetoprotein (AFP), alpha-fetoprotein-L3 (AFP-L3), alanine aminotransferase (ALT), and AFP-L3/AFP ratio levels are potential risk factors for malignization in hepatic cirrhosis patients (RR = 2.99, 2.92, 2.72, and 2.34); serum Golgi protein 73 (GP73) level of hepatic cirrhosis patients decreased significantly after developing HCC (t = 2.212; p = 0.041). The detection of ALT, AFP, AFP-L3, and GP73 has a certain guiding significance to predict the risk of HCC in hepatic cirrhosis patients.

Performance verification and comparison of TianLong automatic hypersensitive hepatitis B virus DNA quantification system with Roche CAP/CTM system.

AIM: To investigate and compare the analytical and clinical performance of TianLong automatic hypersensitive hepatitis B virus (HBV) DNA quantification system and Roche CAP/CTM system. METHODS: Two hundred blood samples for HBV DNA testing, HBV-DNA negative samples and high-titer HBV-DNA mixture samples were collected and prepared. National standard materials for serum HBV and a worldwide HBV DNA panel were employed for performance verification. The analytical performance, such as limit of detection, limit of quantification, accuracy, precision, reproducibility, linearity, genotype coverage and cross-contamination, was determined using the TianLong automatic hypersensitive HBV DNA quantification system (TL system). Correlation and Bland-Altman plot analyses were carried out to compare the clinical performance of the TL system assay and the CAP/CTM system. RESULTS: The detection limit of the TL system was 10 IU/mL, and its limit of quantification was 30 IU/mL. The differences between the expected and tested concentrations of the national standards were less than ± 0.4 Log10 IU/mL, which showed high accuracy of the system. Results of the precision, reproducibility and linearity tests showed that the multiple test coefficient of variation (CV) of the same sample was less than 5% for 102-106 IU/mL; and for 30-108 IU/mL, the linear correlation coefficient r2 = 0.99. The TL system detected HBV DNA (A-H) genotypes and there was no cross-contamination during the "checkerboard" test. When compared with the CAP/CTM assay, the two assays showed 100% consistency in both negative and positive sample results (15 negative samples and 185 positive samples). No statistical differences between the two assays in the HBV DNA quantification values were observed (P > 0.05). Correlation analysis indicated a significant correlation between the two assays, r2 = 0.9774. The Bland-Altman plot analysis showed that 98.9% of the positive data were within the 95% acceptable range, and the maximum difference was -0.49. CONCLUSION: The TL system has good analytical performance, and exhibits good agreement with the CAP/CTM system in clinical performance.

Age and Ebola viral load correlate with mortality and survival time in 288 Ebola virus disease patients.

BACKGROUND: A Chinese medical team managed Ebola virus disease (EVD) patients in Sierra Leone from October 2014 to March 2015 and attended to 693 suspected patients, of whom 288 had confirmed disease. METHODS: A retrospective study was conducted of the 288 patients with confirmed disease. Clinical symptoms, manifestations, and serum viral load were analyzed and compared among the different groups for mortality and survival time. RESULTS: Among the 288 confirmed EVD patients (149 male and 139 female, median age 28 years, and median log viral load 6.68), 98 died, 36 recovered, and 154 were lost to follow-up. Common symptoms were fever (77.78%), fatigue (64.93%), abdominal pain (64.58%), headache (62.85%), and diarrhea (61.81%). Compared to patients aged<18 years, those who were older than 40 years had a higher probability of death (odds ratio 2.855, p=0.044). Patients with a viral load of >10(6) copies/ml had a higher case fatality rate than those with <10(6) copies/ml (odds ratio 3.095, p=0.004). Cox regression showed that age, viral load, and the presence of diarrhea correlated with mortality. CONCLUSION: Patients with a high viral load, of older age, and with diarrhea had a higher mortality and shorter survival time.

NOTCH reprograms mitochondrial metabolism for proinflammatory macrophage activation.

Metabolic reprogramming is implicated in macrophage activation, but the underlying mechanisms are poorly understood. Here, we demonstrate that the NOTCH1 pathway dictates activation of M1 phenotypes in isolated mouse hepatic macrophages (HMacs) and in a murine macrophage cell line by coupling transcriptional upregulation of M1 genes with metabolic upregulation of mitochondrial oxidative phosphorylation and ROS (mtROS) to augment induction of M1 genes. Enhanced mitochondrial glucose oxidation was achieved by increased recruitment of the NOTCH1 intracellular domain (NICD1) to nuclear and mitochondrial genes that encode respiratory chain components and by NOTCH-dependent induction of pyruvate dehydrogenase phosphatase 1 (Pdp1) expression, pyruvate dehydrogenase activity, and glucose flux to the TCA cycle. As such, inhibition of the NOTCH pathway or Pdp1 knockdown abrogated glucose oxidation, mtROS, and M1 gene expression. Conditional NOTCH1 deficiency in the myeloid lineage attenuated HMac M1 activation and inflammation in a murine model of alcoholic steatohepatitis and markedly reduced lethality following endotoxin-mediated fulminant hepatitis in mice. In vivo monocyte tracking further demonstrated the requirement of NOTCH1 for the migration of blood monocytes into the liver and subsequent M1 differentiation. Together, these results reveal that NOTCH1 promotes reprogramming of mitochondrial metabolism for M1 macrophage activation.

[Changes of lymphocyte subsets in peripheral blood of patients with hepatitis C].

OBJECTIVE: To investigate the changes of lymphocyte subsets in the peripheral blood of patients with hepatitis C. METHODS: A total of 219 patients with hepatitis C and 66 healthy individuals served as normal controls were enrolled in the study. The proportions of T-lymphocytes (CD45(+)CD3(+), CD45(+)CD3(+)CD4(+), CD45(+)CD3(+)CD8(+)), B-lymphocytes (CD45(+)CD3(-)CD19(+)), NK-lymphocyte (CD45(+)CD3(-)CD16(+)56(+)) in peripheral blood were detected by flow cytometry. RESULTS: The count of peripheral lymphocyte subsets of patients with hepatitis C decreased from the stage of chronic infection to decompensated cirrhosis. The numbers of CD4(+), CD8(+)and NK lymphocytes of patients with chronic hepatitis C were significantly lower than those in healthy individuals (P<0.05). In the early and decompensated stage of hepatic cirrhosis, the numbers of lymphocyte subsets of patients were significantly lower than those in patients with chronic hepatitis C (P<0.01), and they were significantly lower in decompensated cirrhosis than in early cirrhosis (P<0.01). The proportion of B lymphocyte and CD4/CD8 ratio of patients increased from chronic infection to decompensated cirrhosis. Compared with chronic infection, the proportion of B lymphocyte and CD4/CD8 ratio in early and decompensated cirrhosis increased significantly (P<0.01 or P<0.05), while the proportion of NK lymphocyte was significantly reduced (P<0.01). CONCLUSION: From the stage of chronic infection to decompensated cirrhosis, patients with hepatitis C were found with the decreased numbers of CD4(+)T, CD8(+) T and NK lymphocytes, and the increased proportion of B lymphocyte and CD4/CD8 ratio.

Matrix-assisted laser desorption ionization: time of flight mass spectrometry-identified models for detection of ESBL-producing bacterial strains.

BACKGROUND: The increase in the amount of extended spectrum beta-lactamases (ESBL)-producing gram-negative bacteria is seriously threatening human health in recent years. Therefore, it is necessary to develop a rapid and reliable method for identification of ESBLs. The purpose of this study was to establish a novel method to discriminate between ESBL-producing and non- ESBL-producing bacteria by using the matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) technique. MATERIAL/METHODS: We detected hydrolyzed production of cefotaxime after incubation with 69 gram-negative bacteria by using MALDI-TOF-MS. Then we established genetic algorithm (GA), supervised neural networks (SNN), and quick classifier (QC) models using several peaks to identify ESBL-producing strains. To confirm the clinical applicability of the models established, a blinded validation test was performed in 34 clinical isolated strains. RESULTS: Using ClinPro Tools software, we identified 4 peaks (456 Da, 396 Da, 370 Da, and 371 Da) in mass spectra of cefotaxime solution that have high enough specificity to discriminate ESBL-producing from non- ESBL-producing strains. Recognition capability of models established were 97.5% (GA), 92.5% (SNN), and 92.5% (QC), and cross validation rates were 90.15% (GA), 97.62 (SNN), and 97.62% (QC). The accuracy rates of the blinded validation test were 82.4% (GA), 88.2% (SNN), and 82.4% (QC). CONCLUSIONS: Our results demonstrate that identification of ESBLs strains by MALDI-TOF-MS has potential clinical value and could be widely used in the future as a routine test in clinical microbiology laboratories.

[Distribution of genotypes in ESBLs producing E. coli strains isolated from posthepatitic cirrhosis' patients with bloodstream infection].

OBJECTIVE: To study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection. METHODS: E. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast. RESULTS: A total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79. CONCLUSION: CTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.

[Establishment of a quantitative detection method for Golgi protein73].

OBJECTIVE: To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum. METHODS: A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. RESULTS: The linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%. CONCLUSION: Established ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.