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

ObjectiveTo investigate the clinical application of tacrulimus (TAC, FK506) in children with primary nephrotic syndrome (NS). MethodsSixty-five primary NS children received routine or decreased-dosage glucocorticosteroid according to clinical NS types after hospitalization. At the same time, TAC was given orally with the dosage of 0.1 to 0.15 mg/kg, once every 12 hours, for 6 to 24 months. And the serum concentration of TAC was monitored during the course. ResultsAfter the treatment of TAC for 1 to 2 months, 65 patients were recovered with gradually reduced urinary protein, rapidly increased serum albumin, and improvement of cholesterol and triglycerides. Total remission rate was 83.1% and onset time was 7 to 54 days. Twelve cases experienced recurrence. Increased CD4, as well as 3/3 or 3/1 TAC genotype, indicated higher remission rate. Various pathological types had different remission rates or ratio, which were as follows: minimal change nephropathy (96.4%), mesangial proliferative glomendonephritis (90.0%), membranous nephropathy (2/3), membranous proliferative glomerulonephritis (3/5), focal segmental glomerulosclerosis (4/9). The patients would recover in the course of treatment under the conditions of TAC initial dose as 0.1 to 0.15 mg /kg per 12 hours and controlled serum concentration as 5 to 10 g/L. During the treatment, 12 cases appeared gastrointestinal symptoms, mainly as anorexia, nausea and vomiting, 1 abdominal pain, 2 headache, 1 tremor, 1 paresthesia, 3 insomnia, 4 transient increased Scr, 8 slightly increased NAG, 6 increased C3 and α-2 macroglobulin. The symptoms disappeared within one week or after stopping TAC. ConclusionsTAC is effective in primary NS children, even with abnormal liver function or tuberculosis infection. TAC can also be a substitute to cyclosporine A.

Objective To modify the methods of operation and establishment of cerebral ischemia rat models made by thread occlusion. Method We randomly divided 120 male SD rats into a common group (n = 50), an improvement group (n = 60) and a sham-operated group (n - 10). Rats in the common and improvement groups were made into models using the common and improvement methods separately. All models were evaluated on the basis of physical sign indices and 2,3,5-triphenyltetrazolium chloride (TTC) staining. The TTC staining results were taken as gold standards. Then, we compared the achievement ratios of the two groups, and computed the sensitivity and specificity of every physical sign index based on these standards. The χ~2 or correction χ~2 test was used to compare the ratios. Results (1) The achievement ratio in the improvement group was significantly higher than that in the common group (71.67% vs. 52.00%, P = 0.034). (2) The sensitivity of evaluation for both common and improvement methods was 98.55%. However, the specificity of evaluation for the improvement method was significantly higher than that for the common method (100.00% vs. 40.00%, P =0.000). Conclusions The establishment achievement ratio and evaluation correctness of models are obviously elevated by modification of the operation and establishment methods.

OBJECTIVE:To evaluate the rationality and validity of chemical medicine tablet production equipment cleaning pro-cedure. METHODS:Among several chemical medicines prepared by similar production technology as Metoprolol succinate sus-tained-release tablets,Captopril tablets,Isosorbide mononitrate tablet and Metformin hydrochloride tablet,Metoprolol succinate sus-tained-release tablets had strongest toxicity and were included in validation test. The production equipment was cleaned and disinfect-ed according to cleaning procedure. The point which was most difficult to clean could be wiped and sampled by using the cotton swab method. The detection limit and the limit of quantitation of the residue limits were verified as well as the recovery rate of wip-ing,in order to evaluate whether the results meet the requirements. RESULTS:The cotton swab method is adopted to wipe sample and detect the point which is most difficult to clean. The visible foreign body has not been found in each sampling point. The amount of residual drug is <29.75 μg/cotton bud,and microbial limits are <50 CFU/cotton bud,indicating test items are in line with the standard. CONCLUSIONS:The cleaning method can effectively clean the production equipment,and can effectively pre-vent product contamination and cross contamination to ensure the quality,efficacy and safety of the next batch of products.

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 assess the left ventricular systolic asynchronicity in chronic ischemic model with real-time three-dimensional echocardiography (RT-3DE), and to explore the affection of low-dose dobutamine to it. Methods A chronic ischemic model was induced by placing an Ameroid constrictor in the left circumflex(LCX) in swines,then full volume RT-3DE was performed by Philips iE33 with X3-1 probe combining rest and stress(dobutamine stress echocardiography, DSE) every week after LCX constriction.Ten normal pigs before operation served as controls (group A). Examination of all the models post operation were grouped into group B (mild stenosis, LCX stenosis＜50% ), group C (moderate stenosis, LCX stenosis 50%～75%) and group D (severe stenosis, LCX stenosis≥75%) according to the results of coronary angiography. Images were copied to QLAB 5.2 postprocess workstation,and 3DQA software was used to analyze the full volume data sets. The time to the point with minimal systolic volume (Tmsv) in each segment was taken to derive the following indexes of systolic synchrony: the maximum difference of Tmsv (Tmsv-dif) and standard deviation(Tmsv-SD) among various segments and standard index (Tmsv-dif% and Tmsv-SD%), to evaluate left ventricular dyssynchrony. Tmsv3-6 represented the maximum difference of Tmsv between lateral segment and posterior septum (Tmsv3-5: between lateral segment and inferior) in basal level. Results Tmsvl2-Dif%, Tmsv6-Dif%, Tmsv3-6% and Tmsv3-5% under stress condition in group C and D were significantly higher than those at rest;all the data in group D were significantly higher than in group A and B, and in group C higher than group A ( P ＜0.05,0.01 ). Compared with group A,Tmsv6-Dif,Tmsv3-6 and Tmsv3-5 in group B were significantly increased under stress condition,and so did their standardize data under both rest and stress conditions ( P ＜ 0.05, 0. 01 ). Conclusions RT-3DEcombined with DSE could display sensitively the left ventricular asynchrony caused by chronic ischemia,and that will be more significant in lateral wall in LCX stenosis than in normal segments.