Objective To establish a real-time fluorescent quantitative PCR ( FQ-PCR ) method for detection of murine norovirus ( MNV) in laboratory mouse and provide the basis for establishment of a standard detection method for MNV.Methods Specific primers were designed and MNV DNA standards were prepared according to the MNV genome sequences published on NCBI .The specificity, sensitivity, repeatability and stability of the established Q-PCR method were tested.The established Q-PCR method was applied to detect 766 mouse caecum content samples to explore preliminarily the infection status of laboratory mice in Beijing .Results No cross reaction showed in human norovirus and feline calicivirus with the established Q-PCR method.The sensitivity was up to 10 copies/μL.The coefficient of variation ( CV) of intra-assay and inter-assay was less than 2%.There were 301 positive cases detected in the 766 samples of laboratory mice.Conclusions The established FQ-PCR method is accurate and effective with high specificity , sensitivity and repeatabiliy in the quantitative detetion of nucleic acid , and can be applied to rapidly and quantitatively screen MNV in laboratory mice .

Objective To observe the influence of several kinase pathways such as extracellular signal-regulated kinase ( ERK) , and mitogen-activated protein kinase p38 ( p38MAPK) on nuclear factor erythroid 2-related factor 2 (Nrf2) pathway activated by Fructus Schisandrae extracts (FSE) . Methods HepG2 cells were treated by FSE for 24 hours after pretreatment with protein kinase inhibitors for 2 hours. The mRNA expression levels of Nrf2 and downstream target genes heme oxygenase-1 (HO-1), NAD (P) H quinine oxidoreductase 1(NQO1), P-glycoprotein ( P-gp) and multidrug resistance-associated protein 2 ( MRP2) were detected by real-time polymerase chain reaction ( RT-PCR) , and their protein expression levels and Nrf2 nuclear translocation were measured by Western blotting method. Results RT-PCR results showed that the mRNA expression levels of HO-1, NQO1, P-gp and MRP2 activated by FSE in HepG2 cells were inhibited by PD98059, SB203580 and Rottlerin, and the mRNA expression of Nrf2 was suppressed only by SB203580 and Rottlerin. Western blotting results showed that the mRNA expression levels of HO-1 and P-gp activated by SCE in HepG2 cells were inhibited by PD98059, SB203580 and Rottlerin. In addition, the protein expression of Nrf2 in HepG2 cytoplasm was increased by the above three inhibitors, and nuclear translocation of Nrf2 was inhibited by PD98059 and SB203580. Conclusion The mechanism of FSE activating Nrf2 pathway may be associated with the increase of Nrf2 nuclear translocation through the direct phosphorylation of Nrf2 induced by ERK and p38MAPK.

Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; therapy ; Combined Modality Therapy ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; genetics ; pathology ; therapy ; Lymphatic Metastasis ; Neoplasm Staging ; Oligonucleotide Array Sequence Analysis ; Prognosis

OBJECTIVE: Tonsillectomy is one of the most frequently applied operations in the ENT practice. This prospective study compared intraoperative records and postoperative clinical outcomes between adults and children patients following monopolar electrocautery tonsillectomy. METHOD: Forty adult patients and Forty children patients with histories of recurrent tonsillitis or hypertrophic tonsillitis were enrolled. Intraoperative parameters and postoperative outcomes were compared. RESULT: Children tonsillectomy with monopolar electrocautery was significantly faster to perform (P < 0.05), and produced significantly less intraoperative blood loss (P < 0.05), and faster to return to commencement of a regular diet (P < 0.05) than adults. Children tonsillectomy endured less postopera- tive pain within a week (P > 0.05) than adults, but there was no significant difference in pain on the 14th postoperative day in two groups. There was no obvious postoperative hemorrhage in two groups. There was no significant difference in postoperative tonsillar fossa healing and postoperative temperature between the groups. CONCLUSION Children and adults tonsillectomy with monopolar electrocautery had clinical characteristics respectively. Monopolar electrocautery tonsillectomy was safe and operated easily in both two groups.
Adult ; Blood Loss, Surgical ; Child ; Electrocoagulation ; Humans ; Pain, Postoperative ; Postoperative Hemorrhage ; Postoperative Period ; Prospective Studies ; Tonsillectomy ; methods ; Tonsillitis ; surgery ; Wound Healing

Objective To investigate the clinical value of prenatal ultrasonography in diagnosing fetal aortic atresia.Methods Ultrasound images of 9 cases with fetal aortic atresia confirmed by autopsy were retrospectively analysed and summarized.Results Fetal echocardiography showed ascending aorta and aortic arch were thin in seven cases and unclear in two cases.Color Doppler flow imaging showed fetal aorta no blood connection with left ventricle and reverse blood flow in aortic arch among all 9 cases.There were 6 cases associated with hypoplastic left heart syndrome,two cases associated with ventricular septal defect and well-developed left ventricle,one case complicated with complete atrioventricular septal defect.Nine cases were detected at average gestational age (26.0 ± 2.6) weeks,minimum gestational age was only 16 weeks pregnant.Conclusions Prenatal ultrasound diagnosis of fetal aortic atresia has important clinical value.Ascending aorta and aortic arch are thin or unclear,no blood flow connection between fetal aorta and left ventricle and reverse blood flow in aortic arch are major diagnostic evidences.Color Doppler flow imaging is helpful for early diagnosis.

Objective To explore the effect of ectopic overexpression of Smac/DIABLO on the proliferation of pancreatic cancer cell line SW1990,and the sensitization to TRAIL and Gemcitabine induced apoptosis.Methods The Smac/DIABLO gene was transfected into the pancreatic cancer cell line SW1990 with the participation of Lipofectamine 2000 (SW1990/Smac).The cell line transfected with empty vector served as controls (SW1990/neo).The SW1990/neo and SW1990/Smac cells were assigned into the following treatment groups:TRAIL group,Gemcitabine group,TRAIL plus Gemcitabine group,and the control group.The SW1990 cells were treated with TRAIL and Gemcitabine in different concentrations and time.The cell growth inhibition rate (CGIR) was detected by MTT,the rate of apoptosis was measured by flow eytometry,the apoptosis morphous was observed by Heochst 33342 staining.The expressions of apoptosis-associated proteins such as Smas/DIABLO,XIAP,cytochrome C and caspase-3 were detected by Western blot.Results The cell growth of SW1990/Smac was significantly lower than growth of SW1990/ neo.The concentration of TRAIL were 200,500,1000 and 2500 ng/ml respectively.After 24 hours,the CGIR of SW1990/neo and SW1990/Smac were 11.11％,46.03％,67.08％,76.19％ and 22.11％,42.67％,56.63％,67.6％ respectively (P ＜ 0.05).The concentration of Gemcitabine were 10,20,40 and 60 μmol/L respectively.After 24 hours,the CGIR of SW1990/neo and SW1990/Smac were 15.2％,34.6％,55.16％,76.4％ and 22.65％,36.85％,55.11％,79.99％ respectively (P＜0.05).The cells of SW1990/neo and SW1990/Smac were treated by TRAIL(500 ng/ml),Gemcitabine (20 μmol/L) and combination group.The apoptosis rate were 5.64％,15.30％,27.27％ and 20.37％,23.27％,67.30％ (P ＜ 0.05) respectively.In combination group,the expressions of activators of caspase such as Smas/DIABLO,cytochrome C and caspase-3 increased significantly,while the expressions of inhibitor of apoptosis protein XIAP decreased.Conclusions Ectopic expression of Smac/DIABLO could induce the apoptosis of SW1990 cell,inhibit the cell proliferation,and enhence the sensitivity of SW1990 cell to TRAIL and Gemcitabine.The mechanism of apoptosis sensitization effect by Smac/DIABLO was associated with significant up-regulation of Smac/DIABLO,cytochrome C,down-regulation of XIAP,and the activation of caspase-3.

Objective To characterize Hfq-dependent phenotypes in stress response and to dissect Hfq-dependent transcription of virulence genes and stress-responsive genes in Vibrio cholera.Methods The hfq null mutant strain (△hfq) and the complemented mutant strain (△hfq/pUC18-hfq) were constructed from the wild-type Vibrio cholera.Comparisons on the motility,biofilm formation,growth under various oxygen-supplying conditions,outer membrane resistance,and sensitivity to oxidative stress were analyzed between the wild type strain and the mutant strains.Reverse-transcript fluorescence quantitative PCR (RT-qPCR) was used to determine the transcriptional levels of target genes in the above mentioned strains.Results △hfq and △hfq/pUC18-hfq strains were successfully constructed.The motility,outer membrane resistance and sensitivity to oxidative stress were reduced,but biofilm formation was enhanced in the hfq null mutant strain.RT-qPCR testified that Hfq had regulation effects on gene transcription for forming falagellum,extracellular polysaccharide,outer membrane protein and oxidative stress in Vibrio cholera.Conclusion As a RNA chaperone,Hfq could affect Vibrio cholera in its biofilm formation,resistance to oxidative stress and antibiotics resistance through regulating the transcription of multiple metabolic genes and virulence genes,which indicates that Hfq,combined with other regulators,may play a key role in the complex regulation of metabolic genes and virulence genes.

OBJECTIVE: To study the characteristics of coronary arteriography and traditional Chinese medicine syndrome of 1,069 patients with coronary artery disease (CAD). METHODS: One thousand and sixty-nine patients with CAD were investigated by epidemiological method. The patients were divided into young patients (n=82, aged 45 years or younger) and middle-aged and old patients (n=987, older than 45 years). The characteristics of the two groups were analyzed, including clinical data, coronary arteriography and traditional Chinese medicine syndrome. RESULTS: Compared with middle-aged and old patients, proportion of male, triglyceride, total cholesterol, smoking patients, acute myocardial infarction and family history of CAD in young patients were significantly higher (P<0.05). Patients accompanying with hypertension and diabetes in middle-aged and old patients were more than those in young patients (P<0.05). Occurrence rates of morbidity of left circumflex coronary artery, left main coronary artery and multi-branch were higher in middle-aged and old patients (P<0.05), however, the occurrence rates of morbidity of single and double-branch were higher in young patients (P<0.05). The occurrence rates of syndromes of qi stagnation and phlegm turbidity in young patients were higher than those in middle-aged and old patients (P<0.05). But the proportions of cold coagulation, yin deficiency, yang deficiency and kidney deficiency in middle-aged and old patients were obviously higher (P<0.05). CONCLUSION: The traditional Chinese medicine syndrome and pathological changes of CAD in young patients are different from those in old patients.

Objective To explore the pathogenic bacteria species of the patients with lactation acute mammitis and their sensitivities to antibiotics in order to provide a guideline in choosing antibiotics reasonably. Methods Four hundred and thirty-three samples from diseased breast(include milk,puncture fluid and secretion in ulcerateing skin)from 310 patients with lactation acute mammitis,who were treated by the center of mammary gland of Maternal and Child Health Care of Haidian district in BeiJing from Jan. to Aug. in 2012. Statistics analysis was made to analysis the pathogenic bacteria species and the characteristics of drug sensitivity. Results (1)Of the 433 samples,407 strains of pathogens were picked out,which consisted of 215 strains of Staphylococcus aureus(SAU),43 strains of methicillin-resistant staphylococcus aureus( MRSA),43 strains of coagulase negative staphylococcus(CNS),42 strains of Methicillin-resistant staphylococcus epidermis (MRSE),22 strains of Staphylococcus epidermidis(SE),12 strains of Methicillin-resistant coagulase negative staphylococci(MRScon)and 8 strains of Alpha hemolytic streptococcus.(2)There were 237 strains of G +bacteria,and the drug sensitive test showed that they were all sensitive to vancomycin(100% ),91. 8% were sensitive to penicillin,57. 0% to clindamycin( CLDM))and 69. 0% to erythrocin. Meanwhile,94. 0% were sensitive to levofloxacin,70. 0% to cephalosporins,and 80. 0% to gentamicin. In addition,there were 43 MRSA, and drug sensitivity test showed that 95. 0% were sensitive to levofloxacin,96. 0% to gentamicin,97. 0% to macrodantin,and 92. 0% to cotrimoxazole. Meanwhile,it's drug resistance rate to erythrocin was 13. 0% and 15. 0% to clindamycin( CLDM). MRSA was completely resistant to Lactam antibiotics( 100% ) such as penicillin and Cephalosporins,95. 0% to levofloxacin,96. 0% to gentamicin,97. 0% to furadantin and 92. 0% to bactrim. Conclusion The predominant pathogenic bacteria of lactation acute mammitis include SAU,MRSA, MRSA,CNS,MRSE,SE and they are all the G + bacteria. They are showed with a high rate of drug resistance, especially for the sensitive drugs which had been proved in the past. Due attention should be paid to germiculture from diseased breast and drug sensitive test as soon as possible,and to provide a guideline in choosing antibiotics reasonably.