Objective To investigate the expressions of NBS1 mRNA and protein in the salivary gland of irradiated rats and explore the role of NBS1 in the repair of radiation injury of salivary gland epithelial cells.Methods Eighty rats were randomly divided into two groups for radiation and control (n =40 each).The rats were fractionally exposed to 3 Gy of 60Co γ-rays once in two days,leading to an accumulation dose of 3,6,9,12,15 Gy.The sham-irradiated controls were anesthetized in parallel but without irradiation.After 2-4 h of irradiation,the rats were sacrificed,IHC and RT-PCR were used to detect the expressions of NBS1 protein and mRNA in parotid and submandibular glands,and the ultra-structural changes in the glands were observed by a transmission electron microscopy.Results After irradiation,the salivary glands became atrophy and the parotid gland cells were damaged more serious than the submandibular gland cells.Compared with the controls,with the groups of dose,at 9,12,15 Gy in parotid gland (t =7.10,17.93,20.86,P ＜ 0.05),at 12,15 Gy in the submandibular gland (t =3.13,7.53,P ＜0.05),the expression of NBS1 mRNA was reduced.With the groups of dose at 9,12,15 Gy in paretid gland (t =4.29,17.91,91.29,P ＜ 0.05 ),the dose at 12,15 Gy in submandibular gland ( t =4.61,11.84,P＜0.05),the expression of NBS1 protein in serous cells,and the dose at 12,15 Gy in parotid gland ductal epithelial cell ( t =3.09,5.62,P ＜ 0.05) were reduced.But in the ductal epithelial cells as well as muoass cells in the submandibualr gland were steadily.Conclusions After irradiation,NBS1 at both protein and mRNA levels was dropped in the salivary gland of rats,which might contribute to the repair of radiation injury of salivary gland.

In near-infrared (NIR) analysis of plant extracts, excessive background often exists in near-infrared spectra. The detection of active constituents is difficult because of excessive background, and correction of this problem remains difficult. In this work, the orthogonal signal correction (OSC) method was used to correct excessive background. The method was also compared with several classical background correction methods, such as offset correction, multiplicative scatter correction (MSC), standard normal variate (SNV) transformation, de-trending (DT), first derivative, second derivative and wavelet methods. A simulated dataset and a real NIR spectral dataset were used to test the efficiency of different background correction methods. The results showed that OSC is the only effective method for correcting excessive background.
Algorithms ; Artifacts ; Computer Simulation ; Coptis ; chemistry ; Models, Chemical ; Models, Statistical ; Multivariate Analysis ; Plant Extracts ; analysis ; chemistry ; Principal Component Analysis ; Spectrophotometry, Infrared ; methods

OBJECTIVETo study the efficacy of endotracheal lavage in neonatal ventilator-associated pneumonia (VAP).

METHODSFifty-eight neonates with VAP between January 2002 and December 2008 were randomly assigned to two groups: lavage and control (n=29 each). After withdrawal from ventilator, both groups received sensitive antibiotics therapy according to sputum culture results as well as supportive treatment. The lavage group was additionally treated with endotracheal lavage (2-3 times daily). The therapeutic effects were compared between the two groups.

RESULTSThere were no significant differences in the average time of mechanical ventilation between the lavage and the control groups. The effective rate in the lavage group (93%) was significantly higher than that in the control group (69%; p<0.05). Three percent of patients in the lavage group required twice or more mechanical ventilation compared with 24% in the control group (p<0.05). Blood gas analysis results were obviously improved in the lavage group 2 hrs after treatment (p<0.01).

CONCLUSIONSEndotracheal lavage can decrease the number in mechanical ventilation and improve therapeutic effects in neonates with VAP.

Female ; Humans ; Incidence ; Infant, Newborn ; Male ; Pneumonia, Ventilator-Associated ; epidemiology ; therapy ; Respiration, Artificial ; statistics & numerical data ; Therapeutic Irrigation ; methods ; Trachea

OBJECTIVETo study the prevalence of hepatitis B virus (HBV) genotype in 5 cities of Fujian province and the clinical implications of distinct genotypes in HBV-related liver diseases.

METHODSHBV genotype was determined by the restriction fragment length polymorphism analysis in patients with chronic HBV infection in 5 cities of Fujian province. The relationship between HBV genotype and its clinical implications was studied by multinomal logistic regression and correspondence analysis.

RESULTSOf the 431 HBV DNA positive patients detected by PCR, 275 (63.8%) belonged to HBV genotype B, 100 (23.2%) to genotype C, 51 (11.8%) to genotype D and D-mixed genotype. Genotype A, E and F were not found. Multinomal logistic regression showed that genotype B was more prevalent in Quanzhou and Sanming cities than in Fuzhou (P = 0.002, P = 0.006), and genotype B appeared significantly more common in asymptomatic carriers (ASC), chronic hepatitis B (CHB) and severe hepatitis (SH). Genotype C was most prevalent in patients with liver cirrhosis (LC) (47.0%) than in those with ASC (14.5%) and SH (14.7%) (P = 0.009, P < 0.001). The positive rate of hepatitis B e antigen was higher in patients with genotype C than in those with genotype B and genotype D (56.0% vs. 52.4%, P = 0.008, and 56.0% vs. 30.8%, P = 0.051, respectively). By correspondence analysis, genotype D and D-mixed genotype seemed to be correlated with hepatocellular carcinoma (HCC).

CONCLUSIONS(1) The major popular genotypes of HBV were B, C and D in Fujian. (2) Data of our study suggested that the geographic distribution of genotype B and C might be different in some cities of Fujian. (3) Genotype B might have a tendency to lead to SH in younger patients with chronic hepatitis B and the development of LC might be associated with genotype C among the elder patients. (4) Genotype D appeared to associate with development of HCC, which called for further study to confirm.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; Female ; Gene Frequency ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; genetics ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

OBJECTIVEThis study was to clarify E-cadherin expressions in non-small cell lung cancer (NSCLC) and its correlation with patients' prognosis.

METHODSTissue microarrays (TMAs) containing specimens from 365 different NSCLC were constructed, covering all stages and almost all histological types of this disease. Slides were immunohistochemically stained with antibodies against E-cadherin. Expression pattern of the protein was analyzed with relation to the clinicopathological. Correlations of the results with patients' overall survival were also examined.

RESULTSImmunohistochemical staining revealed that E-cadherin protein was localized mainly on membranes and the cytoplasm of NSCLC tumors cells. Reduced E-cadherin expression was evident in 32.1%. Reduced E-cadherin expression significantly correlated with lymph nodes metastasis (chi(2) = 16.430, P = 0.001), histological dedifferentiation (chi(2) = 9.243, P = 0.010) and advanced clinical stage (chi(2) = 9.421, P = 0.024). There was no significant difference in E-cadherin expression between squamous cell carcinoma and adenocarcinoma. E-cadherin reduced expression correlated with a poor prognosis (P < 0.0001) in univariate analysis. Multivariate analysis showed a significantly lower survival probability for patients with reduced E-cadherin (P < 0.001), and E-cadherin was an independent prognostic factor for survival of NSCLC patients.

CONCLUSIONSIt suggests that dysfunction of E-cadherin has an important impact in the progression of lung cancer. As an independent prognostic factor, expression of E-cadherin can predict outcome of different group, together with conventional prognostic factors, and subsequently make appropriate management.

Adult ; Aged ; Aged, 80 and over ; Cadherins ; biosynthesis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; mortality ; secondary ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; mortality ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Survival Rate

Objective To explore whether TNF-α involves in the modulation of Cysteinyl leukotriene receptor 1 (CysLT1) expression in bronchial epithelial cells.Methods The bronchial epithelial cell lines 16HBE cells were stimulated with different concentration (0.00,0.05,0.50,5.00,20.00 μg/L) of TNF-α for 48 hours,and CysLT1 mRNA in 16HBE cells was measured by reverse transcription(RT)-PCR.CysLT1 expression was detected by immunohistochemistry.Results 16HBE cells did not express CysLT1,after the cells were treated with TNF-α,obvious expression of CysLT1 were detected by immunohistochemistry.The weak CysLT1 mRNA expression was observed by RT-PCR in 16HBE cells,and after the cells were treated with TNF-α for 48 hours,CysLT1 mRNA expression were upregulated.When the concentrations of TNF-α were 0.00,0.05,0.50,5.00,and 20.00 μg/L respectively,the relative intensities of CysLT1 mRNA/β-actin were 0.048,0.105,0.177,0.182,0.495,respectively.Conclusions TNF-α can upregulate CysLT1 mRNA expression in 16HBE ceils in a dose-dependent manner.When infected by virus,respiratory tract produces abundant TNF-α.The TNF-α can upregulate the expression of CysLT1 in epithelial cells,enhance inflammation reaction in respiratory tract.This may explain partially the mechanism of exacerbation of asthma induced by respiratory tract infection.

Objective To understand molecular characteristics of Japanese encephalitis virus ( JEV ) isolated from the major Japanese encephalitis epidemic areas in Sichuan Province,and to provide the foundation for JEV prevention.Methods 13 JEV strains were isolated from mosquitoes in Sichuan during 2007-2010,E genes and preM genes were sequenced and phylogenetic analyses were performed using MEGA5 molecular software.Results Phylogeoetic analysis indicated that all 13 JEV strains from Sichuan belonged to genotype Ⅰ,homologies at nucleotide level and deduced amino acid level in PreM gene were 97％-100％ and 98.7％-100％,and 97.8％-99.9％ and 99.6％-100％ in E gene,respectively.Homologies at nucleotide level and deduced amino acid level in PreM gene between 13 JEV strains and JEV isolated in 2004 in Sichuan were 96.2％-99.1％ and 97.5％-98.7％,and were 97.7％-99.6％ and 98.6％-100％ in E gene,respectively.By comparison with vaacine strains P3 and SA14-14-2,homologies at nucleotide level and deduced amino acid level were 84.1％-85.8％ and 93.7％-96.2％ in PreM gene,and were 87.6％-88.3％ and 97％-97.8％ in E gene,respectively.The neurovirulence-related 8 amino acid sites encode by E gene remained unchanged in 13 JEV strains.Conclusion JEV with genotype Ⅰ predominated in Sichuan,nucleotide sequences and deduced amino acid sequences in PreM gene and E gene were highly conserved,key neurovirulence-rerlated sites remained unchanged.It suggested currently used vaccine is still capable of preventing JEV infection.

Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition (EMT) in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the interaction between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and (or) Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. After HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Blotting, Western ; Cadherins ; metabolism ; Cell Line ; Epithelial-Mesenchymal Transition ; Humans ; Immunoprecipitation ; Kidney Tubules, Proximal ; cytology ; drug effects ; metabolism ; Protein Binding ; RNA Interference ; Reverse Transcriptase Polymerase Chain Reaction ; Semaphorins ; genetics ; metabolism ; Transfection ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; metabolism