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Objective: To establish a rapid and specific quantitative real-time PCR (qPCR) method for the detection of SARS-CoV-2 subgenomic nucleocapsid RNA (SgN) in patients with COVID-19 or environmental samples. Methods: The qPCR assay was established by designing specific primers and TaqMan probe based on the SARS-CoV-2 genomic sequence in Global Initiative of Sharing All Influenza Data (GISAID) database. The reaction conditions were optimized by using different annealing temperature, different primers and probe concentrations and the standard curve was established. Further, the specificity, sensitivity and repeatability were also assessed. The established SgN and genomic RNA (gRNA) qPCR assays were both applied to detect 21 environmental samples and 351 clinical samples containing 48 recovered patients. In the specimens with both positive gRNA and positive SgN, 25 specimens were inoculated on cells. Results: The primers and probes of SgN had good specificity for SARS-CoV-2. The minimum detection limit of the preliminarily established qPCR detection method for SgN was 1.5×102 copies/ml, with a coefficient of variation less than 1%. The positive rate of gRNA in 372 samples was 97.04% (361/372). The positive rates of SgN in positive environmental samples and positive clinical samples were 36.84% (7/19) and 49.42% (169/342), respectively. The positive rate and copy number of SgN in Wild strain were lower than those of SARS-CoV-2 Delta strain. Among the 25 SgN positive samples, 12 samples within 5 days of sampling time were all isolated with virus; 13 samples sampled for more than 12 days had no cytopathic effect. Conclusion: A qPCR method for the detection of SARS-CoV-2 SgN has been successfully established. The sensitivity, specificity and repeatability of this method are good.
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Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , RNA Subgenômico , Reação em Cadeia da Polimerase em Tempo Real/métodos , RNA Viral/genética , Sensibilidade e Especificidade , Nucleocapsídeo/química , Teste para COVID-19RESUMO
This study aimed to establish the baseline sensitivity of Botrytis cinerea from Panax ginseng to prochloraz, and ensure the fitness of prochloraz-resistant mutants and the cross-resistance of B. cinerea to prochloraz and commonly used fungicides for the prevention and control of gray mold including boscalid, pyraclostrobin, iprodione, and pyrimethanil. The sensitivity of B. cinerea from P. ginseng to fungicides was determined by the mycelial growth rate method. The prochloraz-resistant mutants were screened out through fungicide domestication and ultraviolet(UV) induction. The fitness of resistant mutants was determined through the stability of subculture, mycelial growth rate, and pathogenicity test. The cross-resistance between prochloraz and the four fungicides was determined by Person correlation analysis. The results showed that all B. cinerea strains tested were sensitive to prochloraz, and the EC_(50) value ranged from 0.004 8 to 0.062 9 μg·mL~(-1), with an average of 0.022 μg·mL~(-1). The sensitivity frequency distribution diagram showed that 89 B. cinerea strains were located within the main peak with a continuous single peak curve, and the average EC_(50) value of 0.018 μg·mL~(-1) was taken as the baseline sensitivity of B. cinerea to prochloraz. The fungicide domestication and UV induction obtained 6 resistant mutants, among which 2 strains were unstable and the other 2 strains showed decreased resistance after multiple generations of culture. Furthermore, the mycelial growth rate and spore yield of all resistant mutants were lower than those of their parents, and the pathogenicity of most mutants was lower than that of their parents. In addition, prochloraz had no obvious cross-resistance with boscalid, pyraclostrobin, iprodione, and pyrimethanil. In conclusion, prochloraz has great potential for controlling gray mold in P. ginseng, and the resistance risk of B. cinerea to prochloraz is low.
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Humanos , Panax , Fungicidas IndustriaisRESUMO
Objective:To evaluate the role of adiponectin in sevoflurane pretreatment-induced improvement in cognitive function in mice with myocardial ischemia/reperfusion (I/R).Methods:Thirty SPF healthy adult male wild-type C57 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using a random number table method: sham operation group (group Sham), myocardial I/R group (group MI/R) and sevoflurane pretreatment group (group SP). Another 10 SPF healthy male adiponectin knockout SPF mice, aged 8-10 weeks, weighing 20-25 g, were selected and served as APNKO group.Myocardial I/R was induced by 30 min occlusion of anterior descending branch of left coronary artery followed by reperfusion.In SP and APNKO groups, sevoflurane pretreatment included 3 cycles of 10-minute inhalation of 2% sevoflurane-93% O 2-5% CO 2 interspersed with 15-minute inhalation of 95% O 2-5% CO 2, and then the model was established.At 1, 2 and 4 days of reperfusion, cognitive function was assessed by Morris water maze test. Results:There was no significant difference in swimming velocity at each time point between the 4 groups ( P>0.05). Compared with group Sham, the escape latency was significantly prolonged, and the frequency of crossing the original platform was decreased at each time point in group MI/R ( P<0.05). Compared with group MI/R, the escape latency was significantly shortened, and the frequency of crossing the original platform was increased at each time point in group SP ( P<0.05). Compared with group SP, the escape latency was significantly prolonged, and the frequency of crossing the original platform was decreased at each time point in group APNKO ( P<0.05). Conclusion:Adiponectin is involved in the process of sevoflurane pretreatment-induced improvement in cognitive function in mice with myocardial I/R.
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Objective:To development a management program of hand function rehabilitation for patients with rheumatoid arthritis. Methods:Based on the International Classification of Functioning, Disability and Health, the primary version of management program of hand function rehabilitation for patients with rheumatoid arthritis was developed after evidence-based research and group discussion. The primary version was consulted to 15 experts twice with Delphi method, and revised as the final program. Results:The enthusiasm of experts was 100% in both rounds, the authority coefficient of experts was 0.90, and the Kendall's W coefficient was 0.204 in the first round and 0.202 in the second round. The final program included an item of evaluation and 19 items of intervention in four dimensions: body function, body structure, activity and participation, and environmental factors. Conclusion:A management program of hand function rehabilitation for patients with rheumatoid arthritis has been prepared for further clinical evaluation.
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OBJECTIVES@#To study the value of serum miR-922 and miR-506 expression levels in the diagnosis and prognostic assessment of childhood acute lymphoblastic leukemia (ALL).@*METHODS@#A total of 132 children with ALL (ALL group) and 80 healthy children (healthy control group) were prospectively selected in this study. Quantitative real-time polymerase chain reaction was used to measure the expression levels of serum miR-922 and miR-506 in both groups. Receiver operating characteristic (ROC) curves were plotted to analyze the diagnostic value of miR-922 and miR-506 for childhood ALL. The Kaplan-Meier method was used to plot survival curves, and multivariate COX regression models were used to analyze the risk factors for poor prognosis in children with ALL.@*RESULTS@#The ALL group had significantly higher expression levels of serum miR-922 and miR-506 than the control group (@*CONCLUSIONS@#The expression levels of miR-922 and miR-506 are of good value in the diagnosis and prognostic assessment of childhood ALL.
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Criança , Humanos , Biomarcadores Tumorais , Estimativa de Kaplan-Meier , MicroRNAs/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Prognóstico , Curva ROCRESUMO
Objective To investigate the clinical impact of neutrophil to lymphocyte ratio (NLR) in the diagnosis of ulcerative colitis.Methods The study included 97 patients with ulcerative colitis (UC) and 95 patients with irritable bowel syndrome (IBS) from June 2009 to June 2016.The differences of NLR between the two groups were analyzed and the result of sensitivity and specificity were calculated.The differencesn of NLR were analyzed between the groups of different severity and in different lesionsseparately.Results NLR were significantly higher in ulcerative colitis than in irritable bowel syndrome (t=2.327,P<0.021).Sensitivity was 69.1 % and specificity was 75.8 %.There was statistic different in groups of different severity determination of newly diagnosed ulcerative colitis (F=8.221,P=0.001)and there was no statistic different between different lesions (F=0.737,P=0.483).Conclusion NLR is valuable in the diagnosis of UC and IBS can determine the degree of inflammation of ulcerative colitis.
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Objective To understand the current status of the sanitation of freshwater aquaculture environments ,and Clo-norchis sinensis infection of freshwater fish in the aquaculture and market in a city of Pearl River Delta region,so as to provide the evidence for formulating the prevention and control strategy of clonorchiasis sinensis. Methods In 2016,based on the dis-tribution of freshwater aquaculture,36 freshwater fish ponds among 14 towns were selected for sampling and investigation,and 10-20 pieces were collected from each pond. Besides,3 aquatic product wholesale markets were included,among which 3-6 stalls were selected from each market,and 20-30 pieces were collected from each stall. The metacercaria in the fish was exam-ined by the digestion method. Results In the 36 fish ponds,there were no toilets with the stool being drained into fish ponds di-rectly,and there was only one pond with duck sheds with the stool being drained into fish ponds directly. Totally 437 pieces of freshwater fish from ponds were detected,with a metacercaria positive rate of 4.35%(19/437). The metacercaria positive fish were distributed in 50%(7/14)of towns and 25%(9/36)ponds. The positive rates of crucian carp,grass carp,dace,aristich-thysnobilis,and tilapia were 13.95%(6/43),4.76%(9/189),4.44(2/45),1.55%(2/129),and 0(0/31)respectively,with statistically significant difference(χ2=13.46,P=0.01). Totally 307 pieces of freshwater fish were collected from the wholesale markets,with a total positive rate of 1.95%(6/307). The positive rate of grass carp and aristichthysnobilis were 3.20%(4/125) and 2.78%(2/72)respectively,and no positive samples were found in crucian carp,dace and tilapia,with no statistically sig-nificant difference among the different fish in the infection rate(Fisher exact P=0.75). Conclusions The sanitation of fresh-water aquaculture environments in a city of Pearl River Delta region is relative good. However,there are different degrees of Clo-norchis sinensis infection of freshwater fish in some aquaculture environments and markets.
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The uterine tetanic contraction and uterine artery blood flow reduction are possible reasons for primary dysmenorrhea (PD). In the present study, we aimed to evaluate the uterine relaxant effect and the influence on uterine artery blood velocity of Ge-Gen Decoction (GGD), a well-known Chinese herbal formula. In female ICR mice, uterine contraction was induced by oxytocin exposure following estradiol benzoate pretreatment, and the uterine artery blood velocity was detected by Doppler ultrasound. Histopathological examination of the uterine tissue samples were performed by H&E staining. Ex vivo studies demonstrated that oxytocin, posterior pituitary, or acetylcholine induced contractions in isolated mouse uterus. GGD inhibited both spontaneous and stimulated contractions. In vivo study demonstrated that GGD significantly reduced oxytocin-induced writhing responses with a maximal inhibition of 87%. Further study demonstrated that GGD normalized oxytocin-induced abnormalities of prostaglandins F2 alpha (PGF2α) and Ca(2+) in mice. In addition, injection of oxytocin induced a decrease in uterine artery blood flow velocity. Pretreatment with GGD reversed the oxytocin response on blood flow velocity. Histopathological examination showed pretreatment with GGD alleviated inflammation and edema in the uterus when compared with the model group. Both ex vivo and in vivo results indicated that GGD possessed a significant spasmolytic effect on uterine tetanic contraction as well as improvement on uterine artery blood velocity which may involve PGF2α and Ca(2+) signaling, suggesting that GGD may have a clinic potential in PD therapy.
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Animais , Feminino , Humanos , Camundongos , Velocidade do Fluxo Sanguíneo , Medicamentos de Ervas Chinesas , Dismenorreia , Tratamento Farmacológico , Camundongos Endogâmicos ICR , Ocitocina , Contração Uterina , ÚteroRESUMO
Objective To explore the influencing factors of insulin resistance among children in the mountainous area. Methods One middle school and 2 primary schools in Longquan city were randomly selected,47 classes of schoolchildren aged 1 0 -1 5 years were selected.Weight,height,waist circumstance (WC),blood pressure,blood glucose (FBG), serum insulin (FINS),triglycerides (TG),high -density lipoprotein (HDL -C)and total cholesterol (TC)were determined.Influencing factors of insulin resistance were analyzed by univariate and multiple logistic regression analysis. Results A total of 2 1 25 students were enrolled in this study.The 75th percentile of HOMA -IR was 2.59 respectively. The univariate analysis showed that HOMA -IR was significantly associated with HDL -C,TG,non HDL -C,BMI,WC, SBP,DBP and FPG (P <0.05).The multiple logistic regression analysis showed that WC (OR =1 .07,95%CI =1 .06 -1 .09),FPG (OR =2.29,95%CI =1 .78 -2.94),HDL -C (OR =0.34,95%CI =0.21 -0.56)and TG (OR =1 .43, 95%CI =1 .1 4 -1 .78)were the risk factors of insulin resistance.Conclusion WC,FPG,TG may be the risk factors of insulin resistance among children in the mountainous area,while HDL -C is the protective factor.
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Objective To find out the resistant situation and drug of Mycobacteria patients in Sichuan and offer foundation for clinical.Methods Two hundred randomized clinical isolates of Mycobacterium were determined by Roche drug sensitivity and minimum inhibitory concentration (MIC) method.Results Of the 200 clinical isolates,192 stains were Mycobacterium tuberculosis(MTB) (96.0%),8 strains (4.0%) were non-tuberculosis mycobacterium(NTM).Of the 192 MTB strains,108( 57.3% ) sensitive strains and 84 (43.7%)stains were resistant to one or more than one drugs.Among these 84 resistant strains 23 were multi-drug resistant ( MDR,12.0% ),4 were extensively drug resistant( XDR,2.1% ).The anti-TB drug resistance rates were:SM(16.7%),INH(20.8%),RFP(17.2%),EMB(10.9%),PI(16.1%),LFX(8.8%),AMK ( 16.7% ),CPM ( 6.2% ),PTA ( 33.3% ),respectively.Conclusion The resistance rate of tuberculosis keeps at a high level in Sichuan,especially the resistance rate of multiple (≥4) drug,we should oar attention.
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<p><b>OBJECTIVE</b>To study the characteristics of soft-tissue integument and the differences between soft-tissue and hard-tissue topography in malocclusions.</p><p><b>METHODS</b>144 female patients, 12-15 years old, were selected. They were divided into class I, class II and class III groups according to the value of angle ANB which was measured on the pre-treatment cephalographs. Each group had 48 patients. Each patient had same type of skeletal pattern and occlusal pattern, full set of permanent teeth and none of cranofacial soft-tissue and hard-tissue diseases. 4 pairs of measurements describing soft-tissue and hard-tissue sagittal facial pattern and the prominence of lips and incisors were measured on each cephalograph. They were angle SnNsB', angle ANB, angle NsSnPos, angleNAPo, UL-SnPos, UI-APo, LL-SnPos and LI-APo. The distribution of soft-tissue sagittal facial pattern in each skeletal group was analyzed. The differences between angle SnNsB' and angle ANB, angle NsSnPos and angle NAPo, UL-SnPos and UI-APo, LL-SnPos and LI-APo were calculated in each patient. Then we calculated the means and the ranges of these differences in each group, the probability of positive and negative difference between each pair of measurements in each group were calculated too. Chi2 test on those probabilities were performed between the three groups. The mean difference between each pair of measurements was then analyzed by ANOVA between the three groups.</p><p><b>RESULTS</b>The disharmony between soft-tissue and hard-tissue sagittal facial pattern was found in 20%-30% of malocclusion patients. There were more or less differences between soft-tissue and hard-tissue topography and the ranges of their variation were quite wide. The soft-tissue integument increasingly tended to augment the convexity of soft-tissue facial profile when skeletal pattern varied from class II to class I to class III, at the same time, tended to increase upper lip prominence and decrease lower lip prominence.</p><p><b>CONCLUSION</b>On the average, soft-tissue integument tends to camouflage the abnormality of hard-tissue topography. But as to individual, the relative independence of soft-tissue integument makes it important to notice the influence of soft tissue on treatment planning and results.</p>
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Feminino , Humanos , Masculino , Cefalometria , Face , Incisivo , Lábio , Má OclusãoRESUMO
<p><b>OBJECTIVE</b>To investigate the microbiological changes of subgingival microbials in patients with gingivitis and wearing fixed orthodontic appliances.</p><p><b>METHODS</b>48 subjects (10 to 17 years old) with gingivitis, and wearing fixed orthodontic appliances, were divided randomly into three groups (placebo, NS and CH). Placebo group had normal saline mouthrinse; only and no oral hygiene instruction (OHI). The NS group had OHI and normal saline mouthrinse; The CH group had OHI and 0.12% chlorhexidine gluconate mouthrinse. Bacterial examinations were carried out on baseline, one week, one month and three months after scaling. The bacterial examination was carried out. The percentage of coccus, bacillus and spirochete was calculated.</p><p><b>RESULTS</b>In placebo group and NS group, the percentage of coccus increased up to the third examination then dropped down. The spirochete's percentage changed inversely. CH group maintained an increasing trend in coccus' percentage and decreasing trend in spirochete's percentage. The percentage changes of coccus and bacillus between placebo group and CH group are statistically significant (P < 0.05).</p><p><b>CONCLUSIONS</b>During the three-month examination, the CH group had better microbiologic change than the other two groups.</p>