Objective:To investigate the specificity and sensitivity of four methods including ultrafastreal-time fluorescence polymerase chain reaction (PCR), real-time fluorescence (RT)-PCR, enzyme-linked immunosorbent assay (ELISA) and gold immunochromatography assay (GICA) for the detection of novel Bunya virus, so as to provide experimental basis for the early diagnosis of severe fever with thrombocytopenia syndrome (SFTS).Methods:Serum samples from 86 clinically diagnosis SFTS patients admitted to the Jinan Infectious Diseases Hospital Affiliated to Shandoug University were tested by ultrafast real-time fluorescence PCR, RT-PCR, ELISA and GICA during June 1 to September 30, 2017. Chi-square test was used for statistical analysis.Results:Among 86 serum samples, the positive rate of novel Bunya virus of ultrafast real-time fluorescence PCR, RT-PCR, IgM-ELISA, IgG-ELISA, IgM-GICA and IgG-GICA were 82(95.34%), 79(91.86%), 41(47.67%), 8(9.3%), 19(22.09%) and 3(3.49%), respectively. The specificity of ultrafast real-time fluorescence PCR was 100%, and the sensitivity was 1×10 3 copies/mL.Repeated amplification test showed that the variation coefficient of the computed tomography value was <2%.During phases one, two and three, the positive rates of ultrafast real-time fluorescence PCR were 41(97.62%), 34(94.44%) and 7(87.50%), and RT-PCR were 39(92.86%), 33(91.67%) and 7(87.50%), respectively. During phases one and two, the positive rate of ultrafast real-time fluorescence PCR was slightly higher. The positive rate of anti-novel Bunya virus antibody (IgM) tested by ELISA had a significant increase from phase one (28.57%)to phase three (87.50%). There were statistical differences between phase two and phase, as well as between phase three and phase one ( χ2=8.347 and 7.561, respectively, both P<0.01). IgM-GICA also had an increase from phase one (14.29%) to phase two (33.33%)( χ2=3.962, P<0.01), while it was still lower than the other tests.In phase one, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG)( χ2=33.740, 55.080, 49.010 and 64.340, respectively, all P<0.01). In phase two, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG) ( χ2=7.700, 46.720, 23.700 and 50.630, respectively, all P<0.01). In phase three, the positive rates of ultrafast real-time fluorescence PCR, RT-PCR and IgM-ELISA were equivalent, which were all higher than those of IgG-ELISA and GICA (both IgM and IgG). The positive rates of RT-PCR and IgG-ELISA, IgM-GICA and IgG-GICA were significantly different (all χ2=6.250, all P<0.05). Conclusion:In the early detection of novel Bunya virus, ultrafast real-time fluorescence PCR has higher sensitivity, specificity, good repeatability and high stability, which greatly reduces the amplification time compared with the traditional RT-PCR, and is of great value in the early and rapid diagnosis of SFTS.

Objective:To investigate the dosimetry advantage of 3D-printed minimally invasive guided template used in local advanced cervical cancer intracavitary combined with interstitial radiotherapy.Methods:A total of 68 cases with locally advanced cervical cancer who were admitted to Hebei Cangzhou Hospital of intergrated traditional Chinese medicine and western medicine from May 2016 to August 2019 were selected. All the patients had eccentric tumor or large tumor (tumor diameter >5 cm) after radiotherapy. Intensity modulated radiotherapy was used for external radiotherapy, and intracavitary combined with interstitial radiotherapy was used for brachytherapy. The prescription dose of high-risk clinical target volume (HR-CTV) is 6 Gy/fraction, once a week, five fractions in total. Sixty-eight patients were randomly divided into two groups, 35 cases in the template group who received minimally invasive 3D printing guided template assisted intrauterine tube implantation and insertion needle implantation, and 33 patients in the free implantation group who received free hand intrauterine tube implantation and insertion needle implantation. The position and depth of the insertion needle were adjusted by CT-guidance, and the final CT image was transmitted to the Oncentra Brachy treatment planning system, then the target volume and organs at risk were delineated for planning and treatment.Results:A total of 340 brchytherapy plans were made, including 175 in the template group and 165 in the free implantation group. The D90 values of the HR-CTV and intermediate-risk clinical target volume (IR-CTV) in the template group were increased ( t=3.63, 2.45, P<0.05), and D2 cm3 values (dose of 2 cm 3 of organ at risk) of bladder, rectum and sigmoid colon were significantly decreased ( t=-2.81, -2.54, -2.33, P<0.05). At the same time, the average CT scanning times of each treatment in the template group was (1.78±0.53) times, the average duration of each treatment was (11.35±3.98) min, and the average number of needles used in each implant treatment was (5.21±1.37). The result of free implantation group was higher than that of the template group. The differences were statistically significant ( t=-2.26, -4.53, -3.21, P<0.05). Conclusions:For localized advanced cervical cancer patients with eccentric or large tumors, the 3D printed minimally invasive guided template for intracavitary and interstitial implantation has obvious dosimetry advantages, and the operation is simpler and the duration is shorter.

The mixed methods research is the third research method following quantitative study and qualitative study which plays an unique role in nursing research. This paper reviewed the application of mixed methods research in nursing research and expounded the design flow of the mixed methods research based on explanatory sequential design as well as the key questions during combining quantitative study with qualitative study.

Objective: To evaluate the efficacy and safety of bevacizumab in the treatment of radiation brain necrosis, and to provide guidance for rational clinical application. Methods: A retrospective analysis was made of 14 patients with radiation brain necrosis who had failed to receive mannitol and hormone treatment or had been treated for the first time. All the patients were divided into two groups according to different treatment regimens (5.0 mg/kg repeated every 14 d and 7.5 mg/kg repeated every 21 d). The changes of enhanced lesions in MRI T1W1 phase and edema in T2W1 phase were compared separately in the two groups. The clinical symptoms, changes of KPS score and adverse drug reactions were recorded. Results: Totally 14 patients completed at least two cycles of treatment. After two cycles of treatment, the KPS scores of both groups were improved. The KPS scores of 5.0 mg/kg group were increased by 31.66 points on average compared with those before treatment. The KPS scores of 7.5 mg/kg group were increased by 27.50 points on average compared with those before treatment. The volume of CRN lesions were decreased(46.0±9.4)%(t=6.57, P<0.05) and the volume of edema lesions decreased(68.9±8.9)%(t=3.32, P<0.05) in 5.0 mg/kg group. In 7.5 mg/kg group, these two parameters were reduced by(53.9±10.7)%(t=7.89, P<0.05 and (77.1±14.3)%(t=4.22, P<0.05), respectively. There was no significant difference in the reduction of the mean volume of CRN lesions and the volume of edema area between the two groups (P>0.05). In this study, the incidence of adverse events in the 5.0 mg/kg group was similar to that in the 7.5 mg/kg group, with no significant difference(P>0.05). Conclusions Bevacizumab can significantly improve the clinical symptoms and quality of life of patients with radiation brain necrosis. The safety of 5.0 mg/kg treatment regimen repeated every 14 d is similar to that of 7.5 mg/kg treatment regimen repeated every 21 d, which can be used for reference in the treatment of CRN.

Background With the development of refractive surgery,phakic intraocular lens implantation (PIOL) for high myopia is proving its outstanding merits in short-term treating outcomes.However,its long-term safety and effectiveness were still in more attention.Objective This study was to evaluate and compare the long-term safety,stability and efficacy of Verisyse PIOL implantation with implantable contact lens (ICL) implantation for high myopia.Methods The clinical data of 18 eyes (9 patients) who received Verisyse iris-claw intraocular lens implantation for high myopia and matched 22 eyes (11 patients) who received ICL implantation for high myopia from 2009 to 2011 were retrospectively analyzed.The patients were followed-up for 2 years after surgery.Uncorrected visual acuity (UCVA),best corrected visual acuity (BCVA),spherical equivalence (SE),axial length,corneal endothelial cell density (ECD),contrast sensitivity (CS),wave-front aberrations and postoperative complications were recorded during the following-up.The efficacy index,predictable and safety index were compared between the two groups.Results There was no significant difference in UCVA,BCVA and SE between Verisyse group and ICL group(t =0.92,1.32,4.32;all at P＞0.05).Non-glare CS under the 1.5,3.0,6.0,12.0 and 18.0 c/d spatial frequencies were insignificantly different between the two groups(Z=0.782,0.956,0.495,0.874,0.293;all at P＞0.05),and the similar outcomes were found in glare CS (Z=0.985,1.254,0.896,1.652,0.492;all at P＞0.05) in postoperative 2 years.Root meas square of total high order aberration (RMSh),vertical-trefoil,vertical-coma,horizontal-coma,spherical aberration(SA),were significantly elevated in the Verisyse group compared with the ICL group (Z =4.72,4.24,3.12,3.65,2.16;all at P ＜ 0.05).The mean efficacy index was 108.49 ± 16.62 in the V erisyse group and 106.71±15.88 in the ICL group,showing a significant difference (t =0.54,P＞0.05).The mean safety index was 140.56±33.89 in the Verisyse group and 143.34±34.56 in the ICL group,with a significant difference between them (t=0.29,P＞0.05).The mean predictable index was-0.25(-0.97,4.23)in the Verisyse group and 0.98 (-1.44,1.52) in the ICL group,and the difference was significant (Z =-2.68,P ＜ 0.05).Conclusions Both Verisyse PIOL implantation and ICL implantation for high myopia are safe,predictable and effective.The vusial quality and predictability of ICL implantation are much better than Verisyse PIOL implantation.A long-term effect should be observed for the further evaluation.

Objective To explore the clinical effect of ulinastatin on capillary injury improvement of renal syndrome hemorrhagic fever and renal leakage. Methods Patients were divided into 2 groups according to the random number table. The experimental group(25 cases)was given with ulinastatin and the control group(25 cases) was given 5% sugar solution as a blank control. All patients were treated with nutritional support ,rehydration , prevention of bleeding and other symptoms. In addition,according to the number of days of fever in the experimental group,the patients were divided into two groups. The experimental group A(9 cases)had fever 1 ~ 4 days;the experimental group B(16 cases)had fever 5 ~ 7 days. All patients were measured in microalbuminuria ,serum creatinine,plasma albumin and other clinical indicators after 7 day treatment. Results Compared with the control group,microalbuminuria of the experimental group was significantly decreased;Creatinine recovery rate was faster than that in the control group;Plasma albumin had significantly increased;The number of symptomatic days of concurrent perfusion of other tissues had also significantly reduced. Compared with the experimental group B , microalbuminuria of the experimental group A was significantly decreased;Creatinine recovery rate was faster than that in the experimental group B;Plasma albumin had significantly increased;The number of symptomatic days of concurrent perfusion of other tissues was also significantly reduced. Conclusion Ulinastatin could effectively treat vascular injury and syndrome due to capillary leakage caused by epidemic hemorrhagic fever virus ,and the best effect occurs in early application in the fever.

OBJECTIVETo investigate the correlation between Shigella flexneri multi-drug resistance and drug resistance gene cassette of integrons.

METHODAll 79 strains of Shigella flexneri were isolated from the feces of children ranged in age from 6 months to 14 years in some hospitals of Jinan, between May 2009 and April 2012.The resistance was detected by Kirby Bauer agar diffusion method, 1, 2 and 3 integron gene was amplified by PCR, the variable region of positive strains treated with enzyme digestion and determined by Series Analysis.

RESULTAmong 79 Shigella flexneri strains, the resistance rate was 91% (72/79) to ampicillin, chloramphenicol, tetracycline, streptomycin, 70% (55/79) to sulfamethoxazole/trimethoprim, 30% (24/79), 23% (18/79), 33% (26/79) and 32% (25/79) to cefotaxime, ceftazidime, ciprofloxacin and levofloxacin.All 79 strains were susceptible to cefoxitin, imipenem, cefoperazone/sulbactam. The common drug resistance pattern is ampicillin tetracycline-chloramphenicol-streptomycin, accounted for 91% (72/79); 91% (72/79) strains carried integrons of class 1, 86% (68/79) strains carried integrons of class 2, No intI3 was detected. The resistance to ampicillin, streptomycin, tetracycline, chloramphenicol of atypical class 1 integron positive strains was significantly higher than the negative strains (χ² = 35.96, P<0.01). The sequencing results:dfrV was detected in class 1 integron variable regions of 9 strains, dfrA17-aadA5 in 2 strains, blaOXA-30-aadA1 in 70 strains, 2 strains were not detected resistance gene cassette, all resistance gene cassettes were dfrA1-sat1-aadA1 in class 2 integron variable regions.

CONCLUSIONThe muti-drug resistance of Shigella flexneri in Jinan was closely associated with integrons.

Adolescent ; Anti-Bacterial Agents ; pharmacology ; Child ; Child, Preschool ; Drug Resistance, Multiple, Bacterial ; genetics ; Dysentery, Bacillary ; microbiology ; Feces ; microbiology ; Humans ; Infant ; Integrons ; Polymerase Chain Reaction ; Shigella flexneri ; drug effects ; genetics

Objective To compare the difference of the positive rates between real-time RT-PCR and ELISA-IgM methods in diagnosis of severe fever with thrombocytopenia syndrome (SFTS) cases.Method 138 serum samples from 78 suspected acute SFTS patients were collected continuously in our hospital,2014.And 91 serum samples from 49 confirmed cases were detected by real-time RT-PCR and ELISA-IgM,simultaneously.The results were analyzed by Software SPSS17.0.Result 138 serum samples were detected by the real-time RT-PCR and ELISA-IgM methods,and the positive rate was 51.45％,43.48％ respectively.There was significant difference between their positive rates (P ＜ 0.05).The courses of disease influenced the positive rates notablely in 49 confirmed cases between the real-time RT-PCR and ELISA-IgM assays.When the serum samples were collected within 8 days onset of illness to be detected,the positive rate of real-time RT-PCR method was higher than that of ELISA-IgM method.While the positive rate of ELISA-IgM showed a rising tendency along with the course extension.Conclusion Real-time RT-PCR would be the first choice for the early diagnosis of suspected SFTS cases.While ELISA-IgM methods could be the necessary supplemental assay and it would be the important diagnostic method as the extension of the course of SFTS.

Objective To analyze the distribution of class 1,2 and 3 integrons and their gene cassettes,and to explore its relationship with drug resistance in Shigella.Methods Antimicrobial susceptibility was detected by minimal inhibitory concentration (MIC) method.All the genes of integrons and gene cassettes were amplified by polymerase chain reaction (PCR).The amplicons were identified by amplified fragment length polymorphism (AFLP) and sequencing.Results Fifty seven multi-drug resistant strains were identified from a total of 62 Shigella strains (91.9％).Among multi drug resistant strains,52 strains carried integrons of class 1 (91.2 ％) and 55 strains carried integrons of class 2 (96.5％).Only 2 strains carried class 1 integrons alone,5 strains carried class 2 integrons alone and 50 strains had both class 1 and class 2 integrons.Class 3 integrons were not detected.The gene cassettes of typical class 1 integrons,dfrV and dfrA17-aadA5,were detected in 6 strains and 2 strains,respectively.Atypical class 1 integrons with gene cassettes blaOxA30 aadA1 were detected in 44 strains.The typical and atypical class 1 integrons coexisted in 6 Shigella flexneri strains.Gene cassettes for class 2 integrons were dfrA1 sat1-aadA1.Conclusions The multi-drug resistant Shigella strains are widely distributed in Ji'nan,and the atypical class 1 integrons and class 2 integrons are common in these strains.Coexistence of the two integrons is observed in some of the strains.

Objective To discuss the characteristics of extended-spectrum beta-lactamases(ESBLs)-producing Shigella and the relation between them and drug-resistance plasmid. Methods The suspicious ESBLs-producing isolates were screened by K-B disc diffusion method, and the ESBLs-producing strains were confirmed by confirmatory test recommended by the National Committee for Clinical Laboratory Standards. Furthermore, the partial blageneof these isolates were detected by PCR using universal primers for TEM, SHV, CTX-M-1 group, CTX-M-2 group and CTX-M-9 group, respectively. The entire blaCTX-M-9 and blaTEM were amplified by PCR using the primers outside the open reading frame (ORF) of these β-1actamases and products were directly sequenced. The conjugation experiment was performed to determine whether the resistance was transferable. Minimal inhibitory concentration (MIC) was detected with double agar dilution method. Results Of the 275 isolates, 12 strains were identified as ESBLs producers. Among them, 8 strains were CTX-M-14 carriers and 4 strains were CTX-M-3 carriers. All ESBLs-producing isolates are positive for plasmid conjugative transfer test. The transconjugants are only resistance to betalactams. Conclusions High resistance to beta-laetams in Shigella is caused by production of ESBLs in the local area. The ESBLs-produeing isolates can transfer the drug resistance through lateral transfer of plasmid.