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 explore the measurement method of the treatment dose of the patient with Diode for photon beam in radiotherapy,and to validate the treatment dose by comparing with the treatment planning system (TPS).Methods Experiments of the reproducibility,dose rate dependence,non-linearity dose response,and calibration factor in 60Co γ and 6 MV X beams were carried out with Diode on the surface of solid phantom and in water phantom.According to the needs of clinic treatment,different conditions were chosen to observe the dose changes with the angle of incidence,energy response,distance of source to skin,field size,wedge angle,block and tray using ionization chamber and water phantom.The Diode was placed on the surface of the solid phantom to obtain the correction factors.The doses of the chest,abdomen,and head and neek were verified with the Alderson phantom and Diode.Diode doses of the pelvis,head and neck at 14 points on the patient were measured.Results The Diode was irradiated at the points of the Alderson phantom,such as AP,RL and LL of the pelvis,with and without wedges,RL and LL junction of the neck and chin,with and without mask,the maximum relative deviation of doses was within ± 3% between Diode and TPS.The Diode was placed in different locations on the patient,including chest,abdomen and head and neck.The relative maximum deviation of doses was within ±5% between Diode and TPS.Conclusions The Diode method is reliable for measuring the exposure doses of the patient in radiotherapy.

Objective To assess the value of multi-slice computer tomography enterography (MSCTE) in demonstrating small intestinal diseases.Methods MSCTE was performed with iso-osmotic manitol (2.5%) as oral contrast in 98 patients with various kinds of suspected small intestinal diseases.All patients were inter- viewed about their tolerance of the procedure.Demonstration of features of various kinds of small intestinal dis- eases was analyzed.MSCTE diagnosis of different small intestinal diseases were compared with the final clinical diagnosis.Results The procedure was acceptable by all patients and no obvious complication was found. MSCTE was performed for 2 patients because of the failure of conventional small bowel enteroclysis.CT features of many kinds of diseases such as tumors,Crohn's disease were clearly displayed.The sensitivity of MSCTE was 96.5% (83/86),accuracy 90.8% (89/98).Conclusion MSCTE is a simple,rapid,noninvasive and effective method in evaluating small intestinal diseases.

Objective To evaluate the role and technical advantages of CT in 125Ⅰseed implantation in the treatment of tumors. Methods CT-guided 125Ⅰ seed implantation in 45 cases of cancer patients treated 57 times in 51 lesions,CT were used to evaluate the progress of lesions after 125Ⅰ seed implantation 1,2,6 months. Results All procedures were once successful and the total effective rate was 74.5% without any serious complications. Fifty CT examinations immediately after implantation showed satisfactorily seeds distribution but seven cases less satisfactorily that include particles overlap,the location is too close to the lesion edge,the larger the radiation cold spots,etc. Follow-up CT reexamination in 1,2 and 6 months demonstrated that CT examinations showed satisfactorily seeds distribution but accurate seeds number,which could be solved by combined with MPR images and the positioning piece. Conclusions CT-guided 125Ⅰ seed implantation treatment of tumors was a safe,reliable and effective minimally invasive treatment. CT in the preoperative treatment of seed implants planning,precise intraoperative guidance,quality assurance and efficacy of postoperative assessment had a greater role and application advantages.

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.