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Objective To compare clinical data of the death in different intensive care unit,in order to provide the medical strategies for patients in EICU.Methods The clinical data of lethal cases from January 1,2013 to December 31,2014 in EICU,SICU and MICU of the First Affiliated Hospital of Sun Yat-sen University were compared.EICU (252 cases),SICU (93 cases) and MICU (80 cases) were enrolled.The demographics of each patient,clinical condition such as critical score (APACHE Ⅱ score),length of stay,overall costs,and the patient families' different opinions to the treatment in each ICU were analyzed.The data was analyzed with SPSS 13.0 software,averaged value was presented as mean ± standard and the non-normal distributions were expressed as median (25％,75％).The one-way analysis of variance was followed by the Tukey post hoc test for pairwise comparisons and chi-square test was used for comparison of percentage between two groups.Results Two hundred and fifty-two cases in EICU had gender ration of 148/96 (male/female),92 cases in SICU 68/24,80 cases in MICU 56/24.Ages of the fatal were EICU 72 ± 17 years,SICU 56 ± 17 years,and MICU 63 ± 20 years,respectively.Age of the fatal in EICU was significantly older than that of the SICU (P ＜ 0.01) and the MICU (P ＜ 0.01).APACHE Ⅱscores were 33 ± 8 in EICU,34 ± 10 in SICU,29 ± 10 in MICU,respectively.The severity scores in EICU patients were higher than those in MICU patients and SICU patients (P =0.01 and 0.021).Lengths of stay were 2 days (1,46) in EICU,14 days (1,84) in SICU,12 days (1,77) in MICU,respectively.EICU hospitalization time was significantly shorter than that of SICU (P ＜ 0.01) and the MICU (P ＜ 0.01).Total costs of hospitalization were 9 777 yuan (400,164 126) yuan in EICU,100 628 yuan (13 639,964 783) yuan in SICU,119 463 yuan (5 650,590 903) yuan in MICU,and that in EICU was significantly less than the total cost of hospitalization in SICU (P ＜ 0.01) and in MICU (P ＜ 0.01).The opinion of patient families was proposed to give up treatment associated with 165 dead cases in EICU,18 death cases in SICU and 20 dead cases in MICU,and the rate of discontinuous treatment in EICU patients was significantly greater than that in SICU (P ＜ 0.01) and in MICU (P ＜ 0.01).There were no significant differences in invasive procedures,invasive hemodynamic monitoring,mechanical ventilation,blood purification and deep vein puncture among three groups.The 5 leading causes in EICU were severe sepsis,stroke,sudden cardiac arrest,acute myocardial infarction and advanced malignancy.Conclusions The death of patients were due to advanced age with severe disease,poor prognosis,and the request of patient family members to give up treatment.The 5 leading causes were severe sepsis,stroke,sudden cardiac arrest,acute myocardial infarction and advanced tumors suggesting the establishment of corresponding treatment scheme to be made and preparation of abundant medical resources to be ready.Timely communication with the patients' families and let them participate in end-stage treatment decisions was the best strategies to improve the successful rate of treating severe patients and use EICU resource effectively.
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<p><b>OBJECTIVE</b>To monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods.</p><p><b>METHODS</b>HCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing.</p><p><b>RESULTS</b>HCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10(-4.618)/0.1 ml and a 50% inhibitory concentration (IC50) to GCV of 5.847 µmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance.</p><p><b>CONCLUSIONS</b>Phenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.</p>
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Humans , Antiviral Agents , Pharmacology , Cell Line , Cytomegalovirus , Genetics , Drug Resistance, Viral , Genetics , Ganciclovir , Pharmacology , Genes, Viral , Genotype , Hematopoietic Stem Cell Transplantation , Mutation , Phosphotransferases (Alcohol Group Acceptor) , GeneticsABSTRACT
Objective To research the antiviral activity of artesunate (ART) in vitro fighting against both standard laboratory strains and ganciclovir(GCV)-resistance strains of human cytomegalovims(HCMV) and to explore whether fractionation dosage method can obviously enhance the antiviral effect of ART.Methods 1.Cytotoxicity assay to ART was performed by the use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetry.The 0％ toxic concentration (TC0) were determined,and median cytotoxic concentration (TC50) was calculated with Probit regression method.2.Antiviral activity assays of ART against HCMV:human embryonic lung fibroblast cells (HELs) were infected with standard laboratory strains and GCV-resistance strains of HCMV,respectively,after which virus was removed and overlays of dulbecco's modified eagle medium(MEM) containing different antiviral drugs were added to the wells.All cells were cultured continuously at 37 ℃ in a 50 mL/L CO2 humidified atmosphere for 7-10 days and the cytopathic effect (CPE) was observed under a microscope.When the degree of CPE was clear (+ + +-+ + + +),the values of absorbency at 490 nm of all cell wells were measured by MTT colorimetry.The cell survival rate (CSR)and drug inhibitory rate (IR) for HCMV were calculated.By Probit regression method,the median inhibitory concentration (IC50) of 2 drugs was calculated respectively.3.To explore whether fractionation dosage method could obviously enhance the antiviral effect of ART against HCMV,the experiment was divided into 3 groups and compared with GCV group,respectively:Group 1:ART antiviral compounds were added to cell layers by one dosage.Group 2:Total drug dosage was divided into 3 parts,and each part was added to cell layers once a day for 3 days.Group 3:Total antiviral compounds were divided into 6 and delivery 2 times a day.The values of absorbency at 490 nm of all cell wells were measured by MTT colorimetry.The CSR and viral inhibitory rates were calculated.All data were statistically analyzed by One-Way ANOVA analyzing using SPSS 18.0 statistical software.P value of ＜0.05 was considered to indicate statistical significance.Results 1.Cytotoxicity assay showed that cytotoxicity was not found in the relevant range of ART concentrations under 62.5 μmol/L.TC0 and TC50 value of ART were 62.5 μmol/L and 171.7 μmol/L.2.In concentration of 5 μmol/L,15 μmol/L and 30 μmol/L,ART and GCV could obviously inhibit growth of HCMV AD169 strains.There was no significant difference between them.The value of GCV IC50 was 3.49μmol/L,and the value of ART IC50 was 2.17 μmol/L.Treatment index (TI) of ART was 28.8,and GCV was 716.3.ART could still obviously inhibit growth of HCMV resistant strains,but GCV couldn't.Differences between them were statistically significant.The value of GCV IC50 to HCMV resistant strains was 44.4 μmol/L,and the value of ART IC50 was 2.5 μmol/L.3.Fractionation dosage method (2 times a day) of ART could improve the inhibition rate of virus significantly compared to that used once a day and single dose method.Difference was statistically significant(P ＜ 0.01).GCV delivered as the same method had little different changes in virus suppression ratio(P ＞ 0.05).Conclusions 1.Cytotoxicity was not found in the relevant range of ART concentrations under 62.5 μmol/L.2.ART could obviously inhibit growth of HCMV resistant strains and standard laboratory strains.3.Fractionation dosage method (2 times a day) of ART could improve the inhibition rate of virus significantly compared to that used once a day and single dose method.4.Because the action mode of ART is different from other anti-HCMV drugs,and ART has a high biological activity and fewer side effects,it is expected to become a kind of new antiviral drugs for HCMV infections.
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Background Evidences, from recent studies, suggested that Borna disease virus (BDV) infection might be associated with human neuropsychosis, especially psychiatric disorders including depressive disorder(DD). However, controversy existed about the association between BDV infection and pathogenesis of DD. This study was to explore further whether the infection of Borna disease virus (BDV) is associated with the pathogenesis of depressive disorder (DD).Methods The p 24 fragment of BDV RNA in peripheral blood mononuclear cells (PBMCs) from 60DD patients and 120 healthy volunteers was detected by nested reverse transcriptase polymerase chain reaction (nRT-PCR) combined with fluorescence quantitative polymerase chain reaction (FQPCR). Positive products were cloned and sequenced before being compared with Strain V and strain He/80, from humans and animals.Results The positive rate (5%, 3/60) of BDV p 24 in PBMCs from the DD patients was significantly higher than that (0%, 0/120) from healthy volunteers ( P＜0. 05). The gene sequence for the positive products showed BDV p 24 in PBMCs from DD patients in Chongqing was most homophylic with H1766 strain detected from iii horses (97.68%), with 2 situs mutations (nt 1675 T→C, nt 1678 C→T), and also similar to the standard strain V(96. 51%)and He/80(95.35 %), with basic exchanges limited to T- C and A→G.Conclusions There was BDV infection in the DD patients in China, which indicated that the pathogenesis of DD in human beings in Chongqing might be associated with the infection of BDV.
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ObjectiveTo study the relationship between Borna disease virus (BDV) and the human viral encephalitis.MethodsThe P24 fragment of BDV RNA in peripheral blood mononuclear cells (PBMC) from 59 patients with viral encephalitis diagnosed clinically, and 112 healthy donors were examined by fluorescence quantitative nested reverse transcriptase polymerase chain reaction (FQ-nRT-PCR).ResultsThere were 3 positive of BDV P24 fragment in 59 patients with viral encephalitis, and no positive in blood donors. The positive rate of BDV p24 in PBMC in viral encephalitis (5.08%) was higher significantly than that in blood donors (P