Phosphoproteomic analysis of human umbilical venous endothelial cells with DENV-2 infection / 南方医科大学学报

OBJECTIVE@#To analyze the differentially phosphorylated proteins in DENV-2-infected human umbilical venous endothelial cells (HUVECs) and explore the possible pathogenic mechanism of DENV-2 infection.@*METHODS@#The total proteins were extracted from DENV-2-infected HUVECs and blank control HUVEC using SDT lysis method. The phosphorylated proteins were qualitatively and quantitatively analyzed using tandem mass spectrometry (TMT). The identified differentially phosphorylated proteins were analyzed by bioinformatics analyses such as subcellular localization analysis, GO enrichment analysis, KEGG pathway analysis and protein-protein interaction (PPI) analysis. Western blotting was used to detect the expressions of phosphorylated Jun, map2k2 and AKT1 proteins in DENV-2-infected HUVECs.@*RESULTS@#A total of 2918 modified peptides on 1385 different proteins were detected, and among them 1346 were significantly upregulated (FC > 1.2, P < 0.05) and 1572 were significantly downregulated (FC < 0.83, P < 0.05). A total of 49 phosphorylated conserved motifs were obtained by amino acid conservative motif analysis. The most abundant differentially phosphorylated peptides in protein domain analysis included RNA recognition motif, protein kinase domain and PH domain. Subcellular localization analysis showed that the differentially modified peptides were mainly localized in the nucleus and cytoplasm. GO enrichment and KEGG pathway analysis showed that the differential peptides were mainly enriched in the regulation of stimulation response, biosynthesis of small molecules containing nuclear bases, and migration of phagosomes and leukocytes across the endothelium. PPI and KEGG joint analysis showed that the up-regulated and down-regulated differentially phosphorylated proteins were enriched in 15 pathways. In DENV-2-infected HUVECs, Western blotting detected differential expressions of phosphorylated proteins related with the autophagy pathway, namely JUN, MAP2K2 and AKT1, and among them p-JUN was significantly down-regulated and p-AKT1 and p-MAP2K2 were significantly upregulated (P < 0.01).@*CONCLUSION@#DENV-2 infected HUVECs show numerous differentially expressed proteins. The downregulation of p-JUN and upregulation of p-MAP2K2 and p-AKT1 suggest their potential roles in regulating autophagy, which is probably involved in the mechanism of DENV-2 infection.

Prognostic analysis of children with Philadelphia chromosome-like acute lymphoblastic leukemia common genes / 中华儿科杂志

Objective: To summarize the clinical data and prognosis of children with Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) common genes. Methods: This was a retrospective cohort study.Clinical data of 56 children with Ph-like ALL common gene cases (Ph-like ALL positive group) treated from January 2017 to January 2022 in the First Affiliated Hospital of Zhengzhou University, Henan Children's Hospital, Henan Cancer's Hospital and Henan Provincial People's Hospital were collected, 69 children with other high-risk B cell acute lymphoblastic leukemia (B-ALL) at the same time and the same age were selected as the negative group. The clinical characteristics and prognosis of two groups were analyzed retrospectively. Comparisons between groups were performed using Mann-Whitney U test and χ2 test. Kaplan-Meier method was used for survival curve, Log-Rank test was used for univariate analysis, and the Cox regression model was used for multivariate prognosis analysis. Results: Among 56 Ph-like ALL positive patients, there were 30 males and 26 females, and 15 cases were over 10 years old. There were 69 patients in Ph-like ALL negative group. Compared with the negative group, the children in positive group were older (6.4 (4.2, 11.2) vs. 4.7 (2.8, 8.4) years), and hyperleukocytosis (≥50×109/L) was more common (25% (14/56) vs. 9% (6/69)), the differences were statistically significant (both P<0.05). In the Ph-like ALL positive group, 32 cases were positive for IK6 (1 case was co-expressed with IK6 and EBF1-PDGFRB), 24 cases were IK6-negative, of which 9 cases were CRLF2 positive (including 2 cases with P2RY8-CRLF2, 7 cases with CRLF2 high expression), 5 cases were PDGFRB rearrangement, 4 cases were ABL1 rearrangement, 4 cases were JAK2 rearrangement, 1 case was ABL2 rearrangement and 1 case was EPOR rearrangement. The follow-up time of Ph-like ALL positive group was 22 (12, 40) months, and 32 (20, 45) months for negative group. The 3-year overall survival (OS) rate of positive group was significantly lower than the negative group ((72±7) % vs. (86±5) %, χ2=4.59, P<0.05). Compared with the 24 IK6-negative patients, the 3-year event free survival (EFS) rate of 32 IK6 positive patients was higher, the difference was statistically significant ((88±9) % vs. (65±14) %, χ2=5.37, P<0.05). Multivariate Cox regression analysis showed that the bone marrow minimal residual disease (MRD) not turning negative at the end of first induction (HR=4.12, 95%CI 1.13-15.03) independent prognostic risk factor for patient with Ph-like ALL common genes. Conclusions: Children with Ph-like ALL common genes were older than other high-risk B-ALL patients at diagnosis, with high white blood cells and lower survival rate. The bone marrow MRD not turning negative at the end of first induction were independent prognostic risk factor for children with Ph-like ALL common gene.

Analysis of gene expression profile and construction of ceRNA regulatory network after DENV-2 infection with HUVECs / 西安交通大学学报(医学版)

【Objective】 To establish a co-expression lncRNA-mRNA ceRNA network and explore the potential molecular mechanism of lncRNA in dengue fever. 【Methods】 DENV-2-infected and normal pHUVEC were sequenced and screened for differentially expressed lncRNA and mRNA by gene microarray technology. Differentially expressed mRNA was analyzed by protein-protein interaction (PPI), and significantly related co-expressed lncRNA-mRNA was screened by Pearson’s correlation coefficient. The microRNA (miRNA) that bound to co-expressed lncRNA-mRNA was predicted by the database. The ceRNA network of co-expressed lncRNA-mRNA was constructed by Cytoscape software. Finally differentially expressed mRNAs and co-expressed lncRNA-mRNA were analyzed by GO and KEGG enrichment, and co-expressed lncRNA-mRNA was verified by RT-qPCR. 【Results】 At 48 h and 72 h after infection, 105 and 51 differentially expressed mRNAs were obtained, respectively, while 59 and 29 differentially expressed lncRNAs were obtained, respectively. Furthermore, at the two time intervals, there were 10 differential mRNAs and 5 differential lncRNAs, respectively. PPI analysis of differential mRNAs showed that isocratic values of interleukin 6 (IL6), interferon-induced protein with tetratricopeptide repeats 2 (IFIT2), and 2’-5’-oligoadenylate synthetase 2 (OAS2) were relatively high. The pairing results of lncRNA-mRNA co-expression analysis with the highest correlation coefficients at 48 h and 72 h after infection were XLOC_001966-SMTNL1 and XLOC_001966-ESR2, respectively. According to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, the functions of differentially expressed mRNA and co-expressed lncRNA-mRNA were mainly involved in virus epidemic prevention response, immune response, and signal transduction, as well as the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway, type I interferon, and cytokine receptor interaction. RT-qPCR revealed that lncRNA XLOC-I2-8991 was upregulated in the co-expressed lncRNA-mRNA, whereas all the other lncRNA and mRNA were downregulated. 【Conclusion】 This study initially revealed the potential lncRNA-mRNA co-expression network during dengue virus infection, and found that co-expressed lncRNA-mRNA was mainly enriched in the immune regulation and signal transduction pathways during virus infection. The findings will help further exploration into the infection mechanism of DENV-2.

Effects of occupational mercury exposure on the immune function of workers / 中国职业医学

@#Objective To investigate the effects of mercury on T lymphocytes and serum immune indexes of workers with Methods occupational mercury exposure. A total of 45 workers with occupational mercury exposure were selected as the ， mercury exposure group and 47 workers without occupational mercury exposure were selected as the control group using the judgment sampling method. Cold atomic absorption spectrometry was used to detect the urinary mercury level of the two groups. （ ） +， + +， + + - + Flow cytometry was used to detect the proportion of cluster of differentiation CD 3 CD3CD4 CD3CD8 and CD3CD19 ， - （ - ） - （ - ） cells in peripheral blood and the levels of tumor necrosis factor α TNF α and interleukin 8 IL 8 in serum. The levels of （ ） ， Results immunoglobulin Ig A IgG and IgM in serum were measured by immune nephelometry. The urinary mercury level of （ ： vs ，P ） individuals in the mercury exposed group was higher than that of the control group median 92.7 13.2 μg/g Cr <0.01 . The +， + +， - + proportion of CD3 CD3CD4 CD3CD19 cells in peripheral blood and serum IgG level in the mercury exposed group （ P ）， - - （ P ） decreased all <0.05 and the serum TNF α and IL 8 levels increased all <0.01 compared with the control group. Urinary - + mercury level was negatively correlated with the proportion of CD3CD19 cells in peripheral blood and serum IgG level in the ［ （r） ， ， P ］， study subjects Spearman correlation coefficient S were −0.21 and −0.31 respectively all <0.05 and positively - - （r ， ， P ） ， correlated with serum TNF α and IL 8 levels S were 0.36 and 0.39 respectively all <0.05 . However the urinary mercury （ P ）， +， + +， level was neither correlated with IgA and IgM levels in serum all >0.05 nor with the proportion of CD3 CD3CD4 + + （ P ） Conclusion CD3CD8 cells in peripheral blood all >0.05 . Occupational exposure to mercury can lead to abnormal ， changes in peripheral blood T lymphocyte subsets B lymphocytes and serum immune factors in workers. The mercury load of occupational mercury exposure workers may impact their immune function.

Development and validation of multilayer perceptual neural network in glomerular filtration rate evaluation / 中华肾脏病杂志

Objective:To develop a neural network model for the evaluation of glomerular filtration rate (GFR) based on multilayer perceptual neural network, and to compare with the improved Chinese based creatinine GFR evaluation formula (C-GFR cr) and the evaluation formula (EPI-GFR cr) of the American Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) for the clinical applicability of multilayer perceptual neural network model in evaluating GFR. Methods:A total of 684 chronic kidney disease (CKD) patients used for developing a modified version of China′s based creatinine GFR evaluation formula were taken as the research object. The data of 454 patients were randomly selected as the development group and the data of the other 230 patients were as the verification group. The multilayer perceptual neural network GFR evaluation model (M-GFR cr) was established. With the double plasma GFR as the reference value (rGFR), the correlation, mean difference, mean absolute difference, precision and accuracy of C-GFR cr, EPI-GFR cr and M-GFR cr were compared. Results:Among the 684 CKD patients, there were 352 males and 332 females, with age of (49.9±15.8) years. The correlation between M-GFR cr and rGFR was the highest (Pearson correlation =0.93, P<0.001). The mean difference of M-GFR cr was lower than that of C-GFR cr ( Z=9.929, P<0.001) and EPI-GFR cr ( Z=10.573, P<0.001). The mean absolute difference of M-GFR cr was also lower than that of C-GFR cr ( Z=3.953, P<0.001) and EPI-GFR cr ( Z=4.210, P<0.001). The accuracy of ±15% of M-GFR cr was higher than that of C-GFR cr ( χ2=26.068, P<0.001) and EPI-GFR cr ( χ2=23.154, P<0.001). The accuracy of ±30% of M-GFR cr was also higher than that of C-GFR cr ( χ2=8.264, P=0.001) and EPI-GFR cr ( χ2=11.963, P=0.001). The results of different stages of CKD showed that in the early stage of CKD (CKD 1-2), the mean difference of M-GFR cr was lower than that of C-GFR cr ( Z=7.401, P<0.001) and EPI-GFR cr ( Z=8.096, P<0.001); the mean absolute difference of M-GFR cr was also lower than that of C-GFR cr ( Z=4.723, P<0.001) and EPI-GFR cr ( Z=4.946, P<0.001); the accuracy of ±15% of M-GFR cr was higher than that of C-GFR cr ( χ2=23.547, P<0.001) and EPI-GFR cr ( χ2=26.421, P<0.001); the accuracy of ±30% of M-GFR cr was also higher than that of C-GFR cr ( χ2=12.089, P=0.001) and EPI-GFR cr ( χ2=16.168, P<0.001). But there was no significant difference in the applicability among C-GFR cr, EPI-GFR cr and M-GFR cr in the advanced stages of CKD (CKD 3-5). Conclusion:Compared with the improved Chinese based creatinine GFR evaluation formula C-GFR cr and CKD-EPI evaluation formula EPI-GFR cr, the accuracy of multilayer perceptual neural network model to evaluate GFR in CKD patients has been significantly improved, especially in CKD 1-2 stage.

Macrophage migration inhibitory factor regulates dengue virus type 2-induced autophagy in human umbilical vein endothelial cells / 中华微生物学和免疫学杂志

Objective:To investigate the effects of macrophage migration inhibitory factor (MIF) on AMPK/ERK/mTOR autophagy signaling pathway in primary human umbilical vein endothelial cells (HUVEC) after dengue virus type 2 (DENV2) infection.Methods:The virulence of DENV2 to C6/36 cells was assessed with 50% tissue culture infectious dose (TCID 50). NS1 gene fragments in DENV2-infected HUVEC were detected by RT-PCR. Transmission electron microscopy was used to detect autophagosomes. Western blot was performed to detect the effects of DENV2 infection on the expression of autophagy-related protein LC3-Ⅱ and MIF in HUVEC. The correction of MIF with LC3-Ⅱ was then analyzed. HUVEC were pretreated with MIF inhibitor (ISO-1) or pathway inhibitor (Compound C or U0126), and then the changes in the expression of MIF, adenosine 5′-monophosphate-activated protein kinase (AMPK) pathway-related proteins and LC3-Ⅱ after DENV2 infection were detected by Western blot to reveal the correlation between MIF and AMPK autophagy pathway. Results:The TCID 50 to C6/36 cells was 10 -9.09/ml in this experiment. NS1 gene fragments were detected in DENV2-infected HUVEC. Autophagosomes or autophagolysosomes were observed in the infected HUVEC and there were differences in autophagy induced by different doses of DENV2. The mRNA levels of MIF and LC3-Ⅱ in HUVEC were positively correlated after DENV2 infection. MIF inhibitor affected AMPK, ERK and LC3-Ⅱ levels, but had no significant influence on MIF expression at protein level. Conclusions:MIF could affect autophagy through regulating the AMPK/ERK/mTOR signaling pathway in HUVEC during DENV2 infection.

Descriptive epidemiological study of hyperkalemia among patients in the emergency department / 中华急诊医学杂志

Objective:To describe the incidence, diagnosis rate, treatment rate and treatment pattern of hyperkalemia, and serum potassium retesting rate among hyperkalemia patients in the emergency department.Methods:Data were derived from Military Data Center for Rational Use of Drugs. Patients who accessed emergency medical services (≥18 years old) with record(s) of serum potassium between 2015 and 2017 were included. The data of laboratory test, diagnosis, and treatment were analyzed. The main outcomes included the incidence of hyperkalemia, the diagnosis rate, the treatment rate, treatment pattern and the 7-day retesting rate.Results:A total of 1 039 245 patients who met the above criteria were included, of whom, 36 615 (3.52%) had at least one hyperkalemia event. Among the emergency patients with chronic kidney disease, heart failure, diabetes mellitus and hypertension, the proportions of patients who experienced hyperkalemia were 47.69%, 29.13%, 21.69% and 10.16%, respectively. The diagnosis rate of emergency hyperkalemia patients was 9.23%. The overall hyperkalemia treatment rate was 42.1%. Insulin + glucose injection was the most commonly used therapy for emergency hyperkalemia patients. The overall serum potassium retesting rate within 7 days was 28.8%.Conclusions:Hyperkaliemia is more common and more severe in patients with chronic kidney disease, heart failure, diabetes and hypertension. The diagnosis rate and retesting rate of hyperkalemia are low, suggesting that the identification and management of hyperkaliemia in emergency patients should be strengthened.

PX-12 Promoting Apoptosis of Multiple Myeloma Cell Line H929 Induced by Bortezomib / 中国实验血液学杂志

OBJECTIVE@#To study the effect of PX-12 on apoptosis of multiple myeloma (MM) cell line induced by bortezomib.@*METHODS@#MM cell line H929 cells were divided into PX-12 group, bortezomib group, combination group, and control group. 5.0 μmol/L PX-12, 20 nmol/L bortezomib, combination of the two drugs, and DMSO were given to the above mentioned group, respectively. After culture for 24, 48, and 72 hours, the changes of cell viability were observed, the MM cell activity was detected by MTT method, and the cell cycle distribution and apoptosis of each group was detected by flow cytometry. The intracellular ROS level was measured by H@*RESULTS@#MTT assay showed that after culture for 72 hours, the activity of H929 cells in PX-12 group (P<0.05) and bortezomib group (P<0.01) was significantly lower than that in the control group, while that in the combination group was decreased most significantly (P<0.01). After culture for 48 hours, cells in G1 phase in PX-12 group was decreased to 40%, while cells in S phase and G@*CONCLUSION@#PX-12 can increase the apoptosis of MM cell line H929 induced by bortezomib, which may be caused by increasing of ROS level.

Pre-transfusional screening and identification of irregular red blood cell antibody in different nationalities, Guizhou, China / 中国输血杂志

【Objective】 To analyze the frequency and profile of irregular antibodies in different ethnic groups through screening and identification of irregular antibodies in 67 552 blood recipients in the Affiliated Hospital of Guizhou Medical University. 【Methods】 Irregular antibody screening was carried out in patients with different ethnic groups from August 1, 2016 to July 31, 2019 by microcolumn gel anti human globulin method, and the irregular antibody specificity were identified by panel cells. 【Results】 1)307 out of 67 552 cases were positive for irregular antibody, with the positive rate at 0.45%(307/67 552). Among them, Chuanqing was 1.27%(6/473), Yi 1.15%(4/348), Buyi 1.03%(10/975), Dong 0.58%(3/514), Han 0.44%(273/62 365), Miao 0.42%(5/1 187) and Tujia 0.34%(2/596), with significant differences among nationalities. Irregular antibody detection: the positive rate of female patients(0.56%, 223/41 359) was higher than that of male patients(0.32%, 84/26 193)(P0.05). The yields of irregular antibodies did not differ by ABO blood groups(P>0.05). 3)The specificity of 307 irregular antibody positive cases involved 7 blood group systems, including Rh system 59.28%(182/307), MNSs system 9.12%(28/307), Kidd system 0.65%(2/307), Duffy system 0.98%(3/307), Lewis system 5.86%(18/307), P system 0.65%(2/307), and Digeo system 0.33%(1/307). In addition, 15.64%(48/307) of autoantibodies, 0.65%(2/307) of cold antibodies and 4.93%(15/307) of unclear antibodies were detected. 4)The distribution of anti-D, anti-C and autoantibodies were statistically significant among the Han, Buyi, Chuanqing, Miao, Yi and Dong nationalities(P0.05). 【Conclusion】 The distribution of irregular antibodies in Guizhou is different by nationalities. Routine screening of irregular antibodies for transfused or pregnant patients can increase the safety and efficacy of blood transfusion. Most of the irregular antibodies detected are Rh blood group system. The exposure to irregular antibodies can be reduced by additional detection of blood group antigen other than RhD for blood recipients and donors, as well as the blood transfusion with matched blood group antigens.

Definition of cut-off value of anti-phospholipase A2 receptor antibody suitable for Chinese patients / 中华肾脏病杂志

Objective:To analyze the antibody level of phospholipase A2 receptor (PLA2R) in Chinese patients with primary membrane nephropathy (PMN) and its correlation with clinical indicators, and to explore more suitable cut-off value for Chinese patients.Methods:All hospitalized patients with renal biopsy at Peking University People's Hospital from January to August 2018 were retrospectively reviewed. According to the primary disease, patients were divided into PMN group (including patients with idiopathic membranous nephropathy and atypical membranous nephropathy of unknown cause) and control group (other pathological types, including secondary membranous nephropathy patients). Their clinical and pathological characteristics were analyzed, and the level of serum PLA2R antibodies was detected using the method of enzyme-linked immunosorbent assay (ELISA). Spearman correlation was used to analyze the correlation between PMN patients' blood anti-PLA2R antibody level and clinical indicators. The risk factors of PMN were analyzed by logistic regression model, and the optimal cut-off value of PMN was analyzed by ROC curve.Results:A total of 354 patients were included in this study, including 114 patients in PMN group and 240 patients in control group. The age of PMN group was (51.7±14.1) years old and the ratio of male to female was 2.2∶1. The median concentration of PLA2R antibody in PMN group was 16.87 RU/ml [inter-quartile range ( IQR) 1.88-57.26], which was significantly higher than that in control group (1.43 RU/ml, IQR 1.20-1.62, P<0.001). In PMN group, the concentration of anti-PLA2R antibody was correlated with the 24-hour urine protein ration ( r=0.278, P=0.003) and urine erythrocyte ( r=0.190, P=0.043), but not with serum albumin ( r=-0.149, P=0.114) and serum creatinine ( r=0.136, P=0.149). The ROC curve showed that the sensitivity of distinguishing PMN from other diseases was 69.3% (95% CI 60.3%-77.0%), the specificity was 92.9%(95% CI 89.0%-95.5%), and the area under the curve was 0.859(95% CI 0.813-0.905) when the cut-off value was set as 2.28 RU/ml, which was significantly better than the cut-off value of 20.00 RU/ml (the sensitivity/specificity was 46.5%/97.5%) and 14.00 RU/ml (the sensitivity/specificity was 53.5%/97.1%). Conclusions:PLA2R antibody is one of the main pathogenic antibodies of PMN. In China, it is recommended to lower its cut-off value to 2.28 RU/ml, which can improve the sensitivity of distinguishing PMN from other pathological types without reducing specificity.

Clinical Analysis of Gene Mutation in Adult Patients with B-ALL and Its Influence on Clinical Prognosis / 中国实验血液学杂志

OBJECTIVE@#To investigate the gene mutation in adult patients with B-ALL and its influence on clinical prognosis.@*METHODS@#Clinical data of 226 adult patients with B-ALL were retrospectively analyzed in the period from August 2011 to February 2018. The incidence of gene mutation in all patients were detected, and the influence of mutation gene on clinical prognosis were estimated. Cox regression model were used to evaluate the independent prognostic factors.@*RESULTS@#208 (92.04%) of 226 patients showed gene mutations, and the median mutation number was 2 (0-8). Among them, 54 cases (23.89%) showed 14 or more mutations. The top five mutation types of all patients were SF1, FAT1, MPL, PTPNII and N-RAS respectively. The median OS and median RFS times of 226 patients were 27.0 (5.5-84.0) months and 22.5 (0-81.0) months respectively. The OS and RFS times of Ph@*CONCLUSION@#Gene mutations are common in all adult B-ALL patients, and the clinical prognosis of patients with JAK and epigenetics-related signaling pathway mutations is worsen, while the WBC level closely relates to the clinical prognosis of the patients.

Infection prevention and control of bedside blood purification treatment in patients with COVID-19 / 北京大学学报(医学版)

OBJECTIVE@#To explore the infection prevention and control strategy of bedside blood purification treatment in corona virus disease 2019 (COVID-19) isolation ward, and to evaluate the effect of infection prevention and control management measures.@*METHODS@#We summarized and analyzed the clinical features, infection status, outcome and infection prevention and control measures of bedside blood purification treatment patients in COVID-19 isolation ward from February 8, 2020 to March 31, 2020, analyzed the COVID-19 cross-infection between the patients and medical staffs, and the blood-borne pathogens cross-infection situation between the patients, and analyzed the effect of bundle prevention and control measures in controlling the occurrence and spread of cross-infection.@*RESULTS@#A total of 101 COVID-19 patients were hospitalized in this COVID-19 isolation ward, of whom 10 patients (9.90%) received bedside blood purification treatment and the blood purification treatment method was continuous hemodialysis filtration (CVVHDF), and the 10 patients received 79 times of blood purification treatment in total. The prevention and control management measures adopted included divisional isolation, patient behavior isolation and patient placement, operator personal protection and hand hygiene, dialysis waste fluid disposal, isolation room air purification, object surfaces, medical devices and medical fabrics dis-infection management. There were no occurrence and spread of COVID-19 in the medical healthcare workers and blood-borne pathogens cross-infection in the patients. And all the twice throat swabs (two sampling interval > 1 day) of the medical staffs in COVID-19 virus nucleic acid test were negative. The 2 suspected COVID-19 patients' throat swab virus nucleic acid test and the COVID-19 IgG, IgM were always both negative, the chest CT showed no viral pneumonia.@*CONCLUSION@#Bedside blood purification treatment in the COVID-19 isolation ward, the occurrence and spread of healthcare associated infection can be effectively controlled through effective infection prevention and control management, including divisional isolation, patient behavior isolation and patient placement, operator personal protection and hand hygiene, dialysis waste fluid disposal, isolation room's air purification, object surfaces, medical devices and medical fabrics disinfection, which can provide experience for diagnosis, treatment and prevention and control of patients in the respiratory infectious disease ward.

Investigation and Analysis of Cognition ，Attitude and Behavior of Part of Drug Practitioners in Shenzhen on Compensation of Adverse Drug Reaction Injury / 中国药房

OBJECTIVE：To investigate th e cognition ，attitude and behavior of part of drug practitioners in Shenzhen on compensation of adverse drug reaction （ADR）injury，and to provide relevant reference for the establishment of ADR injury compensation system. METHODS ：Taking marketing authorization holder （MAH），staff of medical institutions at the second level or above and drug regulatory authorities in the relevant meeting held in Shenzhen in 2018 as objects ，on-the-spot anonymous investigation of “questionnaire star ”was used to collect and analyze the cognition ，attitude（including attitude towards ADR compensation and ADR compensation system ）and behavior of the above-mentioned personnel to ADR compensation. The relevant suggestion was put forward. RESULTS & CONCLUSIONS ：A total of 172 questionnaires were collected （all being valid ）. In term of cognition ，the respondents generally had high awareness of ADR （response rate of ADR concept was 82.0％）. In term of attitude to ADR injury compensation ，100 respondents（58.1％）believed that ADR after medication should be compensated ，and 147 respondents（85.5％）believed that compensation should be made for serious and fatal adverse reactions. In term of attitude to ADR injury compensation system ，respondents thought that ADR damage compensation funds be undertaken by MAH for 131 case times （76.2％）. As for the difficulties in the implementation of ADR compensation ，it was considered that the evaluation of the correlation between adverse reactions and drugs led to the difficulties in the implementation of ADR compensation （143 case times ， 83.1％）；most respondents thought that the first thing to do in the implementation of ADR compensation system was to implement laws and regulations （145 case times ，84.3％）. In term of behavior ，104 respondents（60.5％）had processed costs due to ADR ， most of whom chose self-resolved （85 cases，81.7％）；112 respondents （65.1％） chose to inform medical personnels or pharmacies directly. It is suggested that the relevant departments in Shenzhen should explore and establish ADR injury compensation mechanism in Shenzhen by establishing a public oriented platform and mechanism for ADR science popularization （setting up relevant departments ，supervising the public propaganda of news media ，expanding the public propaganda to the commu nity），exploring and establishing ADR injury compensation mechanism （setting up a special legislative pilot of ADR injury regulations ，setting up ADR injury appraisal committee ），and increasing the investment in ADR testing （fully using of “Internet+”Big Data ，allocating special funds ）.

Arterial stiffness and its impact on prognosis in patients with stage 3-5 chronic kidney disease(pre-dialysis) / 中国实用内科杂志

OBJECTIVE: To assess the impact of arterial stiffness on prognosis in patients with chronic kidney disease(CKD)stages 3-5(pre-dialysis).METHODS: 141 patients suffered from chronic kidney disease(CKD)stages 3-5 pre-dialysis were enrolled in this study between April 2006 and November 2010.Automatic pulse wave velocity(PWV)measuring system was used to examine carotid-femoral pulse wave velocity(CFPWV).According to CFPWV level,we divided the patients into elevated CFPWV group(CFPWV ≥12 m/s)and the normal group(CFPWV<12 m/s).Patients were followed up for the occurrence of cardiovascular event,cardiovascular death and all-cause death.Kaplan-Meier methods were used for survival analysis and Cox's proportional hazard regression model were used to analyze risk factors.RESULTS: Two groups were followed-up(93.72±47.93)months.The incidences of cardiac-cerebral vascular event,cardiac-cerebral vascular death and all-cause death were higher in high CFPWV level groups(62.2%vs.21.6%,56.7%vs.15.7%,64.4%vs.19.6%,P<0.05).The level of CFPWV was higher in patients with cardiac-cerebral vascular event,cardiac-cerebral vascular death and all-cause death than those without those events[(15.31±3.41)m/s vs.(12.08±2.94)m/s,(15.66±3.40)m/s vs.(12.14±2.88)m/s,(15.38±3.38)m/s vs.(11.97±2.87)m/s,P<0.01].Kaplan-Meier curve for overall survivals and cardiac-cerebral vascular event free survivals showed a significant distinct between high and normal CFPWV level groups,suggesting that the incidence of cardiac-cerebral vascular events,cardiac-cerebral vascular mortality and all-cause mortality were significantly higher in high CFPWV level group than in normal CFPWV group(P=0.000).Multivariate Cox regression analysis revealed that increased CFPWV and commencing dialysis were the independent risk factors for cardiac-cerebral vascular event,increased CFPWV and CRP and decreased ALB and commencing dialysis were the independent risk factors for cardiac-cerebral vascular mortality and all-cause mortality(P<0.05).CONCLUSION: The levels of CFPWV in pre-dialysis chronic kidney disease(CKD)stage 3-5 patients increases significantly.The incidence of cardiac-cerebral vascular events,cardiac-cerebral vascular mortality and all-cause mortality are significantly higher in elevated CFPWV group than those of normal group in patients with CKD G3-5.The elevated CFPWV is one of independent risk factors of cardiac-cerebral vascular event,cardiac-cerebral vascular mortality and all-cause mortality in patients with pre-dialysis chronic kidney disease.

Influences of interaction between dengue virus type 2-infected human umbilical vein endothelial cells and macrophages on major inflammatory cytokines / 中华微生物学和免疫学杂志

Objective To study the influences on the production of major inflammatory cytokines after co-culturing macrophages with human umbilical vein endothelial cells ( HUVECs) that were infected with dengue virus type 2 (DENV-2). Methods Density gradient centrifugation was used to isolate periph-eral blood mononuclear cells ( PBMC) from concentrated human leukocytes. Adherent monocytes in culture flasks were obtained and stimulated with macrophage colony-stimulating factor ( M-CSF) to prepare macro-phages. The purity of CD14+CD11b+ cells was measured by flow cytometry. Changes in the expression of NS1 at mRNA level in HUVECs were detected by real-time PCR following DENV-2 infection. DENV-2-in-fected HUVECs were co-culture with macrophages in Transwell chambers. A control group was set up by pre-treating HUVECs with sphingosine-1-phosphate (S1P) type 1 (S1P1)-specific receptor agonist CYM-5442 for 24 h to remove the drug before infection and then co-culturing the infected cells with macrophages. Real-time PCR was used to detect the expression at mRNA level of IL-6 and IL-8 in HUVECs and IL-6, IL-8, TNF-α and IL-1β in macrophages. A double-antibody sandwich ELISA was used to detect the expression of above cytokines in culture supernatants. Results After HUVECs were infected with DENV-2, expression of NS1 gene at mRNA level gradually increased to the peak at 24 h (2. 66±0. 53, P<0. 05) and then de-creased. The purity of macrophages detected by flow cytometry was (89. 16±2. 07) ％. Expression of IL-6 and IL-8 at mRNA level in DENV-2-infected HUVECs was up-regulated. The peak values reached at 24 h of IL-6 and IL-8 expression were 16. 10±0. 17 and 29. 76±0. 58, while the expression levels at 24 h in the un-infected group were 1. 46±0. 67 and 1. 60±0. 54, respectively. Expression of IL-6, IL-8, TNF-αand IL-1βat mRNA level in DENV-2-infected macrophages was increased significantly. The levels of IL-6, IL-8, TNF-αand IL-1β expression at 24 h were 45. 82±3. 72, 52. 34±1. 69 (12 h), 8. 94±1. 75 and 30. 96±1. 44 in the infected macrophages, and 1. 16±0. 22, 1. 15±0. 21, 1. 11±0. 09 and 1. 47±0. 31 in the uninfected group. Expression of these cytokines was decreased at every time points after co-culturing of DENV-2-infec-ted HUVECs with macrophages, but still significantly higher than that in the uninfected group. In the co-cul-ture group with DENV-2 infection, CYM-5442 pretreatment significantly decreased the expression at mRNA level of IL-6 and IL-8 in HUVECs (P<0. 01) and that of IL-6, IL-8, TNF-αand IL-1βin macrophages (P<0. 01). Conclusions DENV-2 could infect primary HUVECs, and then activate macrophages to promote the secretion of large amounts of IL-6, IL-8, TNF-αand IL-1β. Moreover, the activated macrophages could reduce the production of inflammatory cytokines in HUVECs to a certain extent.

Mineral and bone metabolism disorder in an adenine-induced rat model of chronic kidney disease / 中国比较医学杂志

Objective To investigate the characteristic changes of biochemical markers of mineral metabolism, vascular calcification, and renal osteodystrophy in an adenine-induced rat model of chronic kidney disease (CKD). Methods A total of 20 male Sprague Dawley rats (SD rats) were randomly divided into two groups: the normal group fed with a diet without adenine, and the CKD group fed with an adenine-containing diet (7. 5 g/kg) for the first 4 weeks and then a diet without adenine for the following 2 weeks. At the end of the 2nd week, serum biochemical markers were detected. At the end of the 6th week, the SD rats were sacrificed and serum biochemical markers were detected once again. The aortas were collected for pathological examination and detection of vascular calcium and phosphorus contents. Femurs and the fifth lumbar vertebrae were taken for bone mineral density (BMD) measurement and bone histomorphometric analysis. Results At the end of the 2nd and 6th weeks, compared with the normal control group, the levels of serum creatinine, urea nitrogen, phosphorus and parathyroid hormone (PTH) in the CKD group were significantly increased (P＜0. 05 or P＜ 0. 01), and the level of serum calcium was significantly decreased (P＜ 0. 05 or P＜ 0. 01). Medial layer vascular calcification of the aorta occurred in 50％ of the rats in the CKD group, but was not observed in the normal control group. Vascular calcium and phosphorus contents were significantly higher in the CKD group compared with the normal control group (P＜ 0. 05). The BMD of total femur, cortical and trabecular bone tissues of the femur, and the fifth lumbar vertebra was significantly decreased in the CKD group (P＜ 0. 05 or P＜ 0. 01). The histomorphometric analysis showed that both bone resorption and bone formation of the trabecular bone in the CKD group were increased, indicating a high bone turnover status. The volumes of both trabecular and cortical bones of rats in the CKD group were significantly lower than that of the normal control group (P ＜ 0. 05 or P ＜ 0. 01). However, the trabecular bone mineralization was not significantly different between the two groups. Conclusions The adenine-induced rat model of chronic kidney disease (CKD) established in this study shows reduced serum calcium and increased serum phosphorus and PTH, and medial layer vascular calcification of the aorta. With respect to renal osteodystrophy, this model shows a high trabecular bone turnover, normal trabecular bone mineralization, and low bone volume of cortical and trabecular bone, which meets the characteristics of osteitis fibrosa. This model may become a useful tool for future study of chronic kidney disease-mineral and bone disorder (CKD-MBD).

Influences of interaction between DENV-2-infected HUVECs and regulatory T cells on major inflam-matory cytokines / 中华微生物学和免疫学杂志

Objective To investigate the influences on major inflammatory cytokines after co-cul-turing regulatory T cells (Treg) with human umbilical vein endothelial cells ( HUVECs) that were infected with dengue virus type 2 (DENV-2). Methods Peripheral blood mononuclear cells (PBMC) were extrac-ted from concentrated human leukocytes by density gradient centrifugation. Treg cells were sorted by immu-nomagnetic beads. Expression of CD4,CD25 and CD127 molecules on the membrane of Treg cells was detec-ted by flow cytometry to identify the purity of Treg cells. HUVECs pretreated with or without sphingosine-1-phosphate S1P type 1 (S1P1)-specific receptor agonist CYM-5442 for 24 h were first infected with DENV-2 and then co-cultured with Treg cells. Expression of IL-6,IL-8,TNF-α,IL-10 and TGF-β at mRNA level was detected by real-time RT-PCR. Levels of IL-6,IL-8,IL-10 and TGF-β in the culture supernatants were detec-ted by a double-antibody sandwich ELISA. Results The purity of Treg cells was (84. 3±0. 5)%. Expression of NS1 at mRNA level in DENV-2-infected HUVECs first gradually increased and then decreased after reac-hing the peak at 24 h (3. 03±0. 26, P<0. 01). Enhanced expression of IL-6,IL-8 and TNF-α at mRNA level in HUVECs was observed after DENV-2 infection ( P<0. 01). Expression of these cytokines at every time point was decreased after co-culturing DENV-2-infected HUVECs with Treg cells ( P<0. 05),but was still higher than that before infection. CYM-5442 pretreatment decreased the expression of IL-6,IL-8 and TNF-α at mRNA level in DENV-2-infected HUVECs and inhibited the secretion of IL-10 and TGF-β by Treg cells that were co-cultured with DENV-2-infected HUVECs. Conclusion Primary HUVECs infected by DENV-2 can enhance the secretion of IL-10 and TGF-β by Treg cells,and the suppressive cytokines produced by Treg cells can reduce the production of inflammatory cytokines by DENV-2-infected HUVECs.

Prevalence and influencing factors of peritoneal calcification in patients with long peritoneal dialysis duration / 中华肾脏病杂志

Objective To investigate the prevalence and related factors of peritoneal calcification in peritoneal dialysis(PD)patients with long dialysis duration,and to explore the relationship between peritoneal calcification and vascular calcification.Methods This cross-section study enrolled PD patients who had received PD for more than 4 years in Peking University People's Hospital.Peritoneal calcification and abdominal aortic calcification were reviewed by CT scan.Demographic data,clinical characteristics,laboratory data including calcium phosphorus metabolism indexes(Ca,P,ALP and iPTH)and PD adequacy were collected.The influencing factors of peritoneal calcification were analyzed by Logistic regression analysis.The correlation between peritoneal calcification and abdominal aortic calcification were tested by Spearman correlation analysis.SPSS 19.0 was used for statistical analysis.Results(1)Seventy-nine PD patients were enrolled:32 males(40.5％);mean age was(58.7±13.1)years and average PD duration was 77.25(58.00,88.00)months.The major primary diseases were glomerulonephritis(46.8％)and diabetic nephropathy(30.4％).(2)6 patients(7.6％)had CT-detectable peritoneal calcification.77(97.5％)patients were found with various degrees of peritoneal thickening.The prevalence of peritoneal calcification was 7.6％in patients with PD duration more than 4 years,10.3％in patients with PD duration more than 6 years,18.8％in patients with PD duration more than 8 years and 40.0％in patients with PD duration more than 10 years,showing an increasing trend.Compared with non-peritoneal calcification group,the patients in peritoneal calcification group received higher doses of Vitamin D(P ＜ 0.001)and lower triglyceride levels(P=0.041).The patients were divided into two groups according to whether dialysis duration was longer than 9 years,and the proportion of patients with long PD duration in peritoneal calcification group was higher(P=0.013).Logistic regression analysis showed that PD duration,calcium and phosphorus metabolism indexes were not independent risk factors of peritoneal calcification.High vitamin D dose was an independent risk factor for peritoneal calcification(B=2.667,OR=14.394,95％CI 1.655-125.165,P=0.016).(3)74 patients were found with abdominal aortic calcification in different degrees,and the prevalence rate of abdominal aortic calcification was 93.7％.Spearman correlation analysis showed that there was no correlation between peritoneal calcification and vascular calcification(r=0.70,P=0.542).Conclusions The prevalence of peritoneal calcification in long PD duration patients is low.Peritoneal calcification may be associated with high Vitamin D dose and long PD duration.

Peritoneal dialysis-related eosinophilic peritonitis: a case report and literature review / 北京大学学报(医学版)

Peritoneal dialysis (PD)-related peritonitis is recognized as a common complication of peritoneal dialysis. Eosinophilic peritonitis is a rare type of non-infection PD-related peritonitis. Eosinophilic peritonitis in continuous ambulatory peritoneal dialysis (CAPD) patients was first reported in 1967. The cause of eosinophilic peritonitis is obscure, however it may be related to some etiologies: (1) hypersensitivity to PD materials, including catheter or dialysate; (2) bacteria, fungal or mycobacterium tuberculosis infection. Clinical investigations include asymptomatic cloudy PD effluent, fever, abdominal pain and eosinophil count elevate in PD effluent. Eosinophilic peritonitis is usually mild and self-limited. With the development of PD, more eosinophilic peritonitis cases and researches were reported. Here, we report a patient on CAPD with eosinophilic peritonitis. A 71-year-old female patient developed end-stage renal disease for 4 years and underwent CAPD (2 000 mL of 1.5% dialysis solution with four exchanges daily) for 5 months. With a history of unclean food, she was hospitalized for complaints of diarrhea, fever and cloudy peritoneal effluent for 10 days. Dialysis effluent showed an elevated white blood cell (WBC) count of 1 980 cell/mm3, with 60% polymorphonuclear cells. She was diagnosed as PD-related peritonitis, and therapy was initiated with intraperitoneal ceftazidime 1 g once a day and vancomycin 500 mg every other day. She was admitted to the hospital as the symptoms were not relieved. Her peripheral blood cell count showed a total WBC count of 6 940 cells/mm3, 36.8% eosinophil. Her PD effluent analysis showed turbidity, total WBC count of 1 480 cells/mm3, and 83% polymorphonuclear cells. Her dialysate bacteria culture, fungus culture, polymerase chain reaction for Mycobacterium tuberculosis (TB-PCR), acid-fast stain were all negative. On admission day 4, the treatments were changed to levofloxacin 200 mg once a day and vancomycin 500 mg every other day. After two weeks of antibiotics treatment, patient's symptoms were not completely improved and her dialysis effluent remained cloudy. Her blood eosinophil count elevated to 36.8%，eosinophil proportion in PD effluent>90% and PD effluent pathological findings showed eosinophil>90%. Eosinophilic peritonitis was diagnosed and a decision was made to give loratadine daily dose of 10 mg orally. The possible reasons might be the patient's allergy to some components of PD solution or connection systems in the beginning of PD, and this bacterial peritonitis episode, as well as the application of vancomycin, might lead to the fact that eosinophilic peritonitis acutely developed. For there was no improvement in clinical symptoms, loratadine was stopped, and the patient was discharged 18 days later, and received follow-up closely. Two months later, eosinophil count in blood and PD fluid decreased to normal range with no symptom. This case reminds us that in any PD-related peritonitis patient with prolonged symptoms after appropriate antibiotic therapy, and typical clinical symptoms, the diagnosis of eosinophilic peritonitis should be considered. For the count and percentage of eosinophils are not routinely reported in most laboratories, doctors need to contact the department of laboratory and the department of pathology, to confirm the cell count and proportion of eosinophils in dialysis effluent, so as to make the definite diagnosis, which can not only avoid antibiotics overuse, but also avoid antibiotics-induced eosinophilic peritonitis (such as vancomycin).

ITRAQ Coupled with 2DLC-MS/MS to Screen and Identify Speci-fic Bio-markers of T-Cell Acute Lymphoblastic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To screen and identify potential biomarkers specific for T-cell acute lymphoblastic leukemia (T-ALL).</p><p><b>METHODS</b>Sera were collected from 20 newly diagnosed B-cell acute lymphoblastic leukemia (B-ALL) patients and 20 T-ALL patients. Proteins were extracted, purified and digested with trypsin. All specimens were analyzed by isobaric tags for relative and absolute quantification (iTRAQ) and two-dimensional liquid chromatography-tandem mass spectrometry (2DLC-MS/MS) in a data-dependent mode. Enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression of serum soluble L-selectin (sL-selectin).</p><p><b>RESULTS</b>A total of 468 proteins were identified from distinct peptides. Compared with B-ALL group, 31 proteins were significantly differentially up-regulated while 7 proteins were significantly down-regulated in T-ALL group, sL-selectin was the higher up-regulated in these differential expression proteins. The overexpression of sL-selectin in T-ALL was verified by ELISA.</p><p><b>CONCLUSION</b>There are the differentially expressed proteins between T-ALL and B-ALL, and the sL-selectin is specific for T-ALL, which can not only become a new biomarker for the diagnosis and prognosis of T-ALL, but also can be used as a potential target for therapy of this leukemia.</p>