ABSTRACT
Abstract When a SARS-CoV-2 RT-qPCR test is performed, it may determine an indirect measureof viral load called cycle threshold (Ct). Respiratory samples with Ct <25.0 cycles are consideredto contain a high viral load. We aimed to determine whether SARS-CoV-2 Ct at diagnosis couldpredict mortality in patients with hematologic malignancies (lymphomas, leukemias, multiplemyeloma) who contracted COVID-19. We included 35 adults with COVID-19 confirmed by RT-qPCR performed at diagnosis. We evaluated mortality due to COVID-19 rather than mortalitydue to the hematologic neoplasm or all-cause mortality. Twenty-seven (27) patients survivedand 8 died. The global mean Ct was 22.8 cycles with a median of 21.7. Among the survivors,the mean Ct was 24.2, and the median Ct value was 22.9 cycles. In the deceased patients, themean Ct was 18.0 and the median Ct value was 17.0 cycles. Using the Wilcoxon Rank Sum test,we found a significant difference (p = 0.035). SARS-CoV-2 Ct measured in nasal swabs obtainedat diagnosis from patients with hematologic malignancies may be used to predict mortality.
Resumen Cuando se realiza una RT-qPCR para SARS-CoV-2, es posible determinar una medidaindirecta de la carga viral llamada umbral de ciclado (Ct). Las muestras respiratorias con Ct<25,0 ciclos se consideran de alta carga viral. Nos propusimos determinar si el Ct para SARS-CoV-2 al diagnóstico predice la mortalidad en pacientes con neoplasias hematológicas (linfomas,leucemias, mielomas) que contrajeron COVID-19. Incluimos 35 adultos con COVID-19 confirmadopor RT-qPCR al diagnóstico. Evaluamos la mortalidad por COVID-19, no la mortalidad por la neo-plasia hematológica o la mortalidad por cualquier causa. De los 35 pacientes, 27 sobrevivierony 8 fallecieron. El Ct global medio fue 22,8 ciclos con una mediana de 21,7 ciclos. Entre lossobrevivientes, el Ct medio fue 24,2 ciclos con una mediana de 22,9 ciclos. Entre los fallecidos,el Ct medio fue 18,0 y el Ct mediano fue 17,0 ciclos. Empleando la prueba de suma de rangosde Wilcoxon, encontramos una diferencia significative (p = 0,035). En pacientes con neoplasiashematológicas infectados con coronavirus, el Ct de SARS-CoV-2 medido en hisopados nasales almomento del diagnóstico podría ser utilizado para predecir la mortalidad.
ABSTRACT
Objective To discuss the clinical application of minimal residual disease in acute myelogenous leukemia(AML) by wt1 mRNA quantitative combined with multi-parameter flow cytometry (FCM).Methods Real time quantitative polymerase chain reaction (qRT-PCR) method was established for detecting wt1 gene expression level in 35 AML patients.The indexes were detected by different subtypes;And 9 cases of ease and 4 cases of recurrence in patients was followed-up and detected the wt1 level.The multiparameter flow cytometry was used to analyze the minimal residual disease in AML.Results The expression of wt1 gene was significantly higher than that of the control group.Significant difference was found(P<0.05).In the newly diagnosed AMLs, wt1 was the highest in M2 and the lowest in M6.Follow-up of 4 AML patients showed that wt1 gene expression level was markedly decreased after CR, but obviously increased after relapse.The proportion of abnormal myeloid cells in different phases significantly changed by FCM.There was no difference of minimal residual disease in AML by qRT-PCR and multiparameter flow cytometry.The ROC curve was used to analyze the recurrent cases to get the threshold value(3.33%).Conclusion The quantitative analysis of wt1 combined with multi-parameter flow cytometry can be used to monitor minimal residual disease in leukemia patients, assess the treatment efficacy and prognosis, and predict the risk of recurrence.
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Objective To investigate the efficiency of 16S rRNA Real-time reverse transcription PCR technique in the diagnosis of periprosthetic joint infection,and compare its sensitivity and specificity with conventional culture.Methods There were 43 revision cases from July 2013 to December 2015.Synovial fluid collected by puncture preoperatively,tissues from five different parts around the prosthesis collected intra-operatively were cultured by blood plate and BacT/Alert FN respectively.The 16S rRNA in interface membrane was detected by real-time reverse transcription PCR as a marker to diagnose PJI.At the same time,the synovial fluid was routinely bacterial cultured.We compared the sensitivity and specificity of two methods.Results There are 22 THAs and 21 TKAs respectively in 43 cases,23 cases diagnosed prosthetic joint infection and 20 cases diagnosed non prosthetic joint infection.The sensitivity of 16S rRNA Real-time reverse transcription PCR is higher than the conventional bacterial culture (78.2％ vs.47.8％).There was no difference in the specificity and PPV and NPV.For PCR in prosthetic joint infection group,Staphylococcus epidermidis in 5 cases,Staphylococcus aureus in 3 cases,streptococcus in 4 cases,E.coli in 2 cases,Staphylococcus lugdunensis,Pseudomonas aeruginosa,Staphylococcus haemolyticus and Mycoplasma in 1 case respectively.For culture in prosthetic joint infection group,Staphylococcus epidermidis in 5 cases,Staphylococcus aureus in 2 cases,Staphylococcus lugdunensis,Pseudomonas aeruginosa,Staphylococcus haemolyticus and E.coli in 1 case respectively.For non prosthetic joint infection group,PCR and culture are all negative.Conclusion The sensitivity of 16S rRNA Real-time reverse transcription PCR is higher than the conventional bacterial culture.
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Objective To investigate the effect of arsenic trioxide on expression VEGF of lymphoma cells. Methods The VEGF mRNA expression was analysed by by Real-time PCR, and VEGF protein expression in Raji and Jurkat lymphoma cell lines by ELISA. Results ATO can inhibit lymphoma cells by inducing apoptosis. ATO induced lymphoma cell apoptosis was due to time.With the period of ATO effecting on cells goes, the expression of VEGF mRNA and the protein were down-regulated significantly (after 24, 48, 72 h). There were, the VEGF mRNA △△Ct data of treated with ATO, at 12 h, for Raji: 0.75±0.15, 72 h, Jurkat: 1.67±0.13. After 72 h, Raji: 8.95±0.38; Jurkat: 9.09±0.16 (f =3.54, P <0.01; t =3.65, P <0.01). And about the VEGF protein, at 12 h, Raji: 198.38±4.37; Jurkat: 563.11±3.81. After 72 h, Raji: 23.55±2.06; Jurkat: 57.11 ±3.88 (t =2.48, P <0.05; t =2.59, P <0.05). Conclusion ATO can inhibit the proliferation of lymphoma cells by down-regulating the expression of VEGF mRNA and its protein.