Chicken embryo fibroblasts(CEFs)are among the most commonly used cells for the study of interactions between chicken hosts and H5N1 avian influenza virus(AIV).In this study,the expression of eleven housekeeping genes typically used for the normalization of quantitative real-time PCR(QPCR)analysis in mammals were compared in CEFs infected with H5N1 AIV to determine the most reliable reference genes in this system.CEFs cultured from 10-day-old SPF chicken embryos were infected with 100 TCID50 of H5N1 AIV and harvested at 3,12,24 and 30 hours post-infection.The expression levels of the eleven reference genes in infected and uninfected CEFs were determined by real-time PCR.Based on expression stability and expression levels,our data suggest that the ribosomal protein L4(RPL4)and tyrosine 3-monooxygenase tryptophan 5-monooxygenase activation protein zeta polypeptide(YWHAZ)are the best reference genes to use in the study of host cell response to H5N1 AIV infection.However,for the study of replication levels of H5N1 AIV in CEFs,the β-actin gene(ACTB)and the ribosomal protein L4(RPL4)gene are the best references.

Actins ; metabolism ; Adenoma, Pleomorphic ; congenital ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Infant ; Male ; Nasopharyngeal Neoplasms ; congenital ; metabolism ; pathology ; surgery ; Rhabdomyosarcoma ; metabolism ; pathology ; Salivary Gland Neoplasms ; congenital ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

OBJECTIVETo discuss the effect of surgical staging and using craft bone with vancomycin for the treatment of calcaneal fractures.

METHODSFrom January 2006 to December 2012,13 patients with open calcaneal fractures were treated including 9 males and 4 females with an average of 35.2 years old ranging from 23 to 66. All cases were emergency cases. According to Sanders classification of calcaneal fractures, 2 cases were type II, 7 cases were type III, 4 cases were type IV. According to Gustilo-Anderson soft tissue injury classification, 8 cases were type II, 2 cases were type III A, 2 cases were type III B, 1 case were type III C. Firstly a thorough debridement or VSD procedures were applied,secondly calcaneal fracture were treated with open reduction, plate fixation and bone graft complex with antibiotics. Based on clinical examination, radiographic evaluation, and American Foot and Ankle Surgery Society (AOFAS), ankle function were evaluated after operation.

RESULTSOpen wounds were headed after dressing and repairing,, lateral calcaneal wound were healed during the first period. All patients were followed up for 6 to 36 months (means 14.5 months). Fracture healing time was 14 to 20 weeks (means 16.2 weeks). Last follow-up AOFAS ankle-hindfoot score was (80.0 +/- 7.4) ranging from 55 to 95.

CONCLUSIONFor patients with open fractures, through reasonable clinical evaluation, staging operation, using bone graft with antibiotics can reduce the incidence of postoperative wound infection and promote fracture healing.

Adult ; Aged ; Anti-Bacterial Agents ; administration & dosage ; Bone Transplantation ; methods ; Calcaneus ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; Fracture Healing ; Fractures, Open ; surgery ; Humans ; Male ; Middle Aged

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Pediatric ; statistics & numerical data ; Lung Diseases ; epidemiology ; mortality ; physiopathology ; Male ; Prognosis ; Prospective Studies ; Respiratory Function Tests ; Survival Rate

Objective:To explore the efifcacy and safety for radiofrequency catheter ablation (RFCA) of left bundle branch guided by left bundle potential (LBP), X-ray image with EnSiteNavX System in canine model.
Methods:The RFCA of left bundle branch was conducted in 13 dogs. A mapping catheter was positioned in right atrium to record right-sided His-bundle (R-His) potential, and an ablation catheter via right femoral artery was retrograded to left ventriclefor LBP mapping and ablation. Meanwhile, EnSiteNavX System was used to identify R-His, L-His and LBP at the same time. The potential characteristics in dogs with successful ablation were observed, the PR interval, QRS shape and time limit, AH interval, HV interval, the A/V electro-gram ratio in ablationcatheter at before and after ablation were recorded. The procedural time and X-ray exposure time between LBP with X-ray image method and LBP, X-ray image with EnSiteNavX System method were compared.
Results: There were 9/13 dogs received successful left bundle branch ablation, 3 dogs failed and 1 suffered from complete A-V block. At the successful ablation target site, the LBP-V was (17.8 ± 2.6) ms with the range of (13-21) ms, and the A/V electro-gram ratio<1/10. The procedural time and X-ray expose time were signiifcantly decreased in LBP, X-ray image with EnSiteNavX System method than those in LBP with X-ray image method P=0.007 and P<0.001.
Conclusion:Under the LBP, X-ray image with EnSiteNavX System guidance method, left bundle branch could be safely and effectively ablated to establish left bundle branch block (LBBB) model in experimental canine.

Objective To probe into the change regulation before and after treatment every snake bite patients for the routine blood test and serum hs‐CRP .Methods The study objects were selected in the hospitalized patients for clear diagnosis belong to what kind of snakes in the past two years .The indexes of routine blood test and serum hs‐CRP were determined before and after treatment various periods in these patients .The test results were made to statistical analysis according to kind of snakes ,periods and disease condition .Results WBC was obvious .rise before treatment only the viper snake bite patients .WBC was all significant .rise after treatment first day and second day for 5 kinds snake bite patients (P<0 .01) .This index had all reduced trend after treatment fourth day but determined value was still high in contrast to the normal reference scope .RBC and HGB all were normal level and had not obvious change before and after treatment for 5 kinds snake bite patients .PLT was reduced before treatment for the trime‐resurus gramineus bite patients ,before and after treatment for the viper snake bite patients (P<0 .05) .The hs‐CRP content was higher before treatment for the viper snake bite patients and was highest after treatment for the cobra snake bite patients ,the inter‐comparison had significant difference (P<0 .05) .Conclusion 5 kinds snake bite patients before and after treatment not basely ane‐mia symptom .But most of the patients appear the inflammatory response .PLT decrease is more serious for the trimeresurus grami‐neus bite patients ,and for the viper snake bite patients especially .

Objective To analyze the abnormal detectable rates of different kinds of blood test indexes before and after treatment in the patients with snake bite and to probe into the change condition of these indexes after different snake bite .Methods The inpa‐tients with clearly diagnosed what kind of snake bite in the past two years were selected as the research subjects .The multiple blood test indexes were determined before and after treatment in these patients .The abnormal detectable rates of these indexes were ana‐lyzed and compared among various snake bite patients .Results The different kinds of snake bite all could cause the different de‐grees of changes in some detection indexes among partial patients .Specially ,the detectable rates of WBC ,PT ,APTT ,TT ,D‐D ,CK , CK‐MB ,LDH ,Urea ,Cr and Cys‐C increase and PLT and Fbg decrease caused by viper bite were apparently higher than those caused by other kinds of snake bites (P<0 .05) .The detectable rates of CO2 increase and K+ decrease in the coral snake bite were apparently higher than those in trimeresurus gramineus ,cobra and viper bite (P<0 .05) .Conclusion Because the snake species and toxicities are different in the various snake bites ,so the caused changes and the abnormal detectable rates of blood test indexes also are different .

Objective:To investigate the effects of different doses of sodium arsenic (NaAsO 2) on mRNA transcription levels of nucleotide excision repair (NER) related genes in normal hepatocytes (L-02 cells) and the modification levels of histone H3 ninth lysine dimethylization (H3K9me2) in the promoter region. Methods:L-02 cells were treated with 0 (the control group) , 5, 10 and 20 μmol/L NaAsO 2 for 24 h ( n = 3). Single cell gel electrophoresis (SCGE) was used to detect DNA damage [Olive tail distance (OTM) and Tail DNA percentage (Tail DNA%)] in L-02 cells. The mRNA expression levels of Xeroderma pigmentosum (XP) gene A (XPA), XP gene D (XPD) and XP gene F (XPF) were detected by real-time fluorescence quantitative PCR. The modification levels of H3K9me2 in XPA, XPD and XPF gene promoter regions (CHIP1 and CHIP2) were detected by quantitative chromatin immunoprecipitation. Results:OTM and Tail DNA% were positively correlated with arsenic doses (in the control and 5, 10 and 20 μmol/L arsenic exposure groups, the values were 0.35 ± 0.09, 0.56 ± 0.18, 3.18 ± 0.31, 4.52 ± 0.55, 0.72 ± 0.05, 1.34 ± 0.26, 3.93 ± 0.43, 5.47 ± 0.65, respectively, r = 0.927, 0.948, P < 0.05). Compared with the control group, the mRNA expression levels of XPA, XPD and XPF in L-02 cells of 10 and 20 μmol/L arsenic exposure groups were significantly lower ( P < 0.05). Compared with the control group, the enrichment levels of H3K9me2 in XPA, XPD and XPF gene promoter regions (CHIP1 and CHIP2) in L-02 cells of 20 μmol/L arsenic exposure group were significantly higher ( P < 0.05). Conclusion:Arsenic may inhibit the transcription of NER related genes by increasing the enrichment level of H3K9me2 in the promoter regions (CHIP1 and CHIP2) of NER related genes, thereby reduce the DNA damage repair ability of L-02 cells, resulting in the aggravation of DNA damage.

Abstract: Objective　To analyze the nucleic acid detection results of severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) by droplet digital PCR (ddPCR) and compare with the detection results of real-time fluorescence quantitative RT-PCR (qRT-PCR), so as to evaluate the advantages and disadvantages of detection, and to provide data support for optimizing the nucleic acid detection scheme of SARS-CoV-2. Methods　According to the SARS-CoV-2 specific primer probe published by the China Center for Disease Control and Prevention, a ddPCR detection method for SARS-CoV-2 was designed. One sample was selected for sensitivity test after gradient dilution; six respiratory virus nucleic acid positive samples including seasonal H3N2 influenza virus and SARS-CoV-2 positive samples were selected for specificity test; five SARS-CoV-2 positive samples were selected for repeatability test; in addition, 30 positive and 20 negative SARS-CoV-2 samples were selected for multiple clinical samples testing, and the results were analyzed and compared with those of qRT-PCR. Results　The ddPCR method can specifically detect SARS-CoV-2, and directly obtain the original copy number of the sample target gene to achieve accurate quantification; the sensitivity test of gradient dilution positive samples showed that qRT-PCR detected target genes in part of the 10-5 dilution of samples, and no target genes were detected in 10-6 dilution, while ddPCR detected all target genes in both 10-5 and 10-6 dilution of samples. The detection limit of ddPCR was two orders of magnitude higher than that of qRT-PCR, and the sensitivity was higher than that of qRT-PCR; in the comparison of the repeatability test results of the two methods, the coefficient of variation of ddPCR was 1.266%-11.814%, lower than 1.729%-26.174% of qRT PCR, and the repeatability was higher than qRT-PCR; among 50 clinical samples, 30 positive samples of confirmed cases of Coronavirus Disease 2019 (COVID-19) were detected by both methods, SARS-CoV-2 was successfully detected by both methods, and 20 negative samples of COVID-19 were detected by both methods, and the results were negative, with a coincidence rate of 100.00% (50/50). Conclusion　The ddPCR method can accurately quantify SARS-CoV-2 with strong specificity, and its sensitivity and repeatability are higher than those of qRT-PCR, but it also has certain detection limitations and is more suitable for the detection of low load samples. In the actual detection, the two methods can be reasonably combined to improve the detection accuracy.

Objective To evaluate the diagnostic values of MSCT for detecting atherosclerotic plaques on New Zealand rabbits models in comparison with pathologic results. Methods Fifteen New Zealand rabbits were enrolled in this study, including 5 with balloon injury and high-fat diet ( group A), 5 with high-fat diet only (group B) and 5 with regular feed (group C). 16th week late, contrast-enhanced MSCT scan was performed in all rabbits with 16 slice MSCT (16-MSCT) in group A and 64 slice MSCT (64-MSCT) in group B and C. The CT and pathological findings were compared in a double-blind manner. The sensitivities and specificities of 16-MSCT and 64-MSCT for detecting atherosclerotic plaques were evaluated by using Fisher test and x2 test. Results Sixty and seventy-five images on 16-MSCT and 64-MSCT had corresponding pathological slices. The sensitivities for the detection of plaques on 16-MSCT and 64-MSCT were 41.5% (22/53) and 64. 9% (24/37), and spocificities of 85. 7% (6/7) and 89. 5% (34/38), respectively. Conclusions 64-MSCT has a higher sensitivity in the detection of atherosclerotic plaques than 16-MSCT. Both scanners can be used to preclude the diagnosis of atherosclerosis.