To investigate the current status of sleep quality among nurses in tertiary hospitals in China, analyze the correlations of work stress and perceived organizational support with the risk of sleep problems, and further examine the mediating effect of perceived organizational support between work stress and sleep problems in nurses.

Objective:To explore potential predictors of refractory Mycoplasma pneumoniae pneumonia (RMPP) in early stage. Methods:The prospective multicenter study was conducted in Zhejiang, China from May 1 st, 2019 to January 31 st, 2020. A total of 1 428 patients with fever >48 hours to <120 hours were studied. Their clinical data and oral pharyngeal swab samples were collected; Mycoplasma pneumoniae DNA in pharyngeal swab specimens was detected. Patients with positive Mycoplasma pneumoniae DNA results underwent a series of tests, including chest X-ray, complete blood count, C-reactive protein, lactate dehydrogenase (LDH), and procalcitonin. According to the occurrence of RMPP, the patients were divided into two groups, RMPP group and general Mycoplasma pneumoniae pneumonia (GMPP) group. Measurement data between the 2 groups were compared using Mann-Whitney U test. Logistic regression analyses were used to examine the associations between clinical data and RMPP. Receiver operating characteristic (ROC) curves were used to analyse the power of the markers for predicting RMPP. Results:A total of 1 428 patients finished the study, with 801 boys and 627 girls, aged 4.3 (2.7, 6.3) years. Mycoplasma pneumoniae DNA was positive in 534 cases (37.4%), of whom 446 cases (83.5%) were diagnosed with Mycoplasma pneumoniae pneumonia, including 251 boys and 195 girls, aged 5.2 (3.3, 6.9) years. Macrolides-resistant variation was positive in 410 cases (91.9%). Fifty-five cases were with RMPP, 391 cases with GMPP. The peak body temperature before the first visit and LDH levels in RMPP patients were higher than that in GMPP patients (39.6 (39.1, 40.0) vs. 39.2 (38.9, 39.7) ℃, 333 (279, 392) vs. 311 (259, 359) U/L, both P<0.05). Logistic regression showed the prediction probability π=exp (-29.7+0.667×Peak body temperature (℃)+0.004×LDH (U/L))/(1+exp (-29.7+0.667×Peak body temperature (℃)+0.004 × LDH (U/L))), the cut-off value to predict RMPP was 0.12, with a consensus of probability forecast of 0.89, sensitivity of 0.89, and specificity of 0.67; and the area under ROC curve was 0.682 (95% CI 0.593-0.771, P<0.01). Conclusion:In MPP patients with fever over 48 to <120 hours, a prediction probability π of RMPP can be calculated based on the peak body temperature and LDH level before the first visit, which can facilitate early identification of RMPP.

Objective:To establishment an assay for HIV-1 intact proviral DNA assay through droplet digital PCR (ddPCR).Methods:DNA was extracted by culturing 8E5 cells, a Tlymphocyte cell line containing a single copy of integrated HIV-1 provirus. Serial diluting DNA were prepared by amplified 1-fold, 5-fold, 25-fold, 625-fold, 3 125-fold, and 15 625-fold across the HIV-1 Ψ region, env region, and eukaryotic chromosome 10 RPP30 regions, and the linear relationship was calculated and the minimum detection concentration. DNA solution of 5 μl, 3.1 μl, 2.5 μl was added to the ddPCR mixture respectively, with each dilution undergoing two batches of detection, and each was repeated four times. The intra-batch variation coefficient was detected, while the inter-batch variation coefficient was detected by the same DNA amount and different DNA amounts to determine the stability; 8E5 cell was used to detect the intact proviral content in cells.Results:The linear fitting goodness of Ψ region, env region and RPP30 region are R2≥0.999, R2≥0.993, R2≥0.996 in 6 dilutions of DNA, respectively. At a 3 125-fold dilution, the lowest positive droplets were detected in the Ψ region, env region and RPP30 region were 3, 2 and 2, respectively, the detected concentrations were 2.37 copies/μl, 1.21 copies/μl and 1.58 copies/μl. The ddPCR repeatability experimental detecting DNA showed that the Ψ region of the intra-batch variation coefficients ranged from 0.66% to 3.43%, with the inter-batch variation coefficients of the same DNA at 3.19%, 4.3% and 3.45% respectively, and the inter-batch variation coefficients of the different DNA at only 4.35%. The env region of the intra-batch variation coefficients ranged from 0.7% to 3.20%, with the inter-batch variation coefficients of the same DNA at 3.18%, 4.52% and 3.4% respectively, and the inter-batch variation coefficients of the different DNA at only 4.02%. The RPP30 region of the intra-batch variation coefficients ranged from 0.91% to 2.91%, with the inter-batch variation coefficients of the same DNA at 3%, 4.55% and 3.37% respectively, and the inter-batch variation coefficients of the different DNA at only 3.98%. The proportion of 8E5 cells containing defective provirus and the proportion of intact provirus were calculated to be approximately 90% and 45%, respectively. Conclusions:Droplet digital PCR used to detect HIV-1 intact proviral DNA, showed strong stability and provided a technical means for HIV-1 infection reservoir detection.

Objective To discover the host factor interacting with severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)papain-like protease(PLpro)and explore the potential mechanism.Methods The second-generation proximity-dependent biotin identification(BioID2)approach combined with mass spectrometry analysis was used to search for the potential host factors.Immunofluorescence and co-immunoprecipitation(Co-IP)assay were used to verify the interactions between DEAD-box helicase 3(DDX3)and PLpro.The influence of PLpro on DDX3-inhibitor of kappa B kinase ε(IKKε)-TANK-binding kinase 1(TBK1)and DDX3-mitochondrial antiviral signaling protein(MAVS)complexes was also investigated by Co-IP.The effect of PLpro on interferon-β(IFN-β)immune pathway and the protease activity on substrates were studied via luciferase activity assay.Results DDX3 could co-locate and interact with PLpro intracellularly.PLpro might possibly inhibit both the formation of DDX3-MAVS complex and the interactions between DDX3-IKK-ε-TBK1.PLpro could negatively regulate type Ⅰ interferon pathway.Overexpression of DDX3 could lead to a significant increase in the cleavage activity of PLpro/PLP-TM that might be significantly decreased in case of inventions with DDX3 expressions.Conclusion DDX3 may be one of the host factors that interact with SARS-CoV-2 PLpro.PLpro negatively regulates IFN-β immune pathway induced by DDX3,which may provide a favorable immune environment for virus replication.

Objective To evaluate the effect of end-stage renal disease(ESRD)hemodialysis(HD)on the prognosis of elderly patients undergoing drug-eluting stent(DES)implantation.Methods A retrospective trial was conducted on 4219 consecutive patients(＞60 years old)undergoing DES treatment at Guro Hospital of Korea University from March 2003 to July 2021,and those with GFR ≥ 1 5 ml/(min·1.73 m2)but not on dialysis were assigned into the non-ESRD group(n=4102)and the others on dialysis into the ESRD group(n=117).Based on their baseline data,a propensity score matching(PSM)model was established,including 183 cases of the non-ESRD group and 99 cases of the ESRD group.The patients were followed up for 1,3 and 5 years,and the incidences of MACE and other endpoint events were recorded during the period.Kaplan-Meier sur-vival curve was plotted,and PSM and multivariate Cox regression models were employed to verify the endpoint events.Results Before PSM,the elderly ESRD group had obviously larger propor-tions of hypertension,diabetes,peripheral vascular disease and other diagnoses than the elderly non-ESRD group(P＜0.01).Univariate Cox regression analysis showed that ESRD was a risk fac-tor for all-cause mortality in the elderly patients at 1,3 and 5 years after DES implantation(P＜0.05,P＜0.01);a risk factor for MACE at 3 and 5 years after implantation(HR=2.67,95％CI:1.73-4.10,P=0.000;HR=2.65,95％CI:1.79-3.90,P=0.000);and a risk factor for develo-ping TVR,TLR,NTVR and revascularization at 5 years after surgery(P＜0.05,P＜0.01).Kaplan-Meier survival curve showed that the incidence of MACE in the elderly ESRD group was higher than that in the elderly non-ESRD group at 5-year follow-up(Plog rank=0.006).Conclusion ESRD significantly increases the risk of long-term MACE after DES implantation in elderly patients.

ObjectiveTo review the ancient and modern literature of Huanglian Ejiaotang and learn about the historical evolution and clinical application， thereby providing a theoretical basis for the modern application of the classical prescription. MethodLiterature in the Chinese Medical Classics Database was retrieved with "Huanglian Ejiaotang" as the keyword. In China National Knowledge Infrastructure （CNKI） and PubMed， "Huanglian Ejiaotang" in Chinese and English was used as the keyword to retrieve literature. The items and modern clinical application studies related to the prescription， medicine， dosage， syndrome， and treatment of Huanglian Ejiaotang were selected and recorded. The inclusion and exclusion criteria were used to screen out literature. The information about the dynasty， book title， function， and indication was integrated to understand the history， evolution， and clinical application of Huanglian Ejiaotang. ResultFinally， 89 ancient books were included with 111 items. Huanglian Ejiaotang was initially recorded in ZHANG Zhongjing's Treatise on Cold Damage and Miscellaneous Diseases in the Han dynasty. It was composed of five herbs， namely Coptidis Rhizoma， Scutellariae Radix， Paeoniae Radix Alba， Asini Corii Colla， and egg yolk. With the change of historical dynasties， the composition， origin basis， dosage， and preparation method of Huanglian Ejiaotang all changed， but the changes in the processing were not obvious， which was basically consistent with Treatise on Cold Damage and Miscellaneous Diseases. In addition， 48 studies were included to analyze the clinical application of Huanglian Ejiaotang， which was mainly used for insomnia， anxiety， depression， diabetes， and so on. ConclusionAccording to the ancient and modern literature， the origin basis， dosage， processing， decoction， administration， and other content of Huanglian Ejiaotang are consistent with Treatise on Cold Damage and Miscellaneous Diseases. The present clinical application has expanded the usage scale of the ancient record， which promotes the innovation and development of the classic prescription and provides references for later research， development， and accurate application.

OBJECTIVE To investigate the effect and mechanism of dihydroartemisinin(DHA)on lipo-polysaccharide(LPS)-induced acute lung injury(ALI)in mice using whole-genome sequencing.METHODS An ALI mouse model was established via intraperitoneal injection of 10 mg·kg-1 lipopolysaccharide.The mice were divided into normal control group(n=10),model group(n=10)and model+DHA group(n=10).The mice in the model+DHA group were injected intraperitoneally with 20 mg·kg-1 DHA,while those in the normal control group and LPS group were injected intraperitoneally with solvent of DHA,saline containing 1%Tween 80 and 10%Macrogol 400.The mice were executed 24 h after drug administration.The wet and dry weight ratio(W/D)of lung tissue was calculated.Hematoxylin-eosin(HE)staining was used to observe histopathological damage in the lung.Classified counts of inflamma-tory cells in alveolar lavage fluid were performed.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α)in alveolar lavage fluid.Real-time quantitative PCR(RT-qPCR)was used to detect mRNA levels of placenta-specific 8(Plac8),Toll-like receptor 7(TLR7),IL-1β,IL-6 and TNF-αin lung tissue.The whole gene transcriptome was sequenced by RNA transcriptome sequencing(RNA-seq)using the Illumina HiSeq high-throughput sequencing platform before the function and signal pathway of differentially expressed gene mRNA between the groups were enriched and analyzed using GO and KEGG enrichment analysis methods.RESULTS Compared with the model group,the lung W/D values of mice,the pathological damage,inflammatory cells in alveolar lavage fluid,expression levels of IL-1β,IL-6 and TNF-α in alveolar lavage fluid(P<0.01,P<0.01,P<0.01),and the mRNA expression levels of IL-1β,IL-6 and TNF-α were significantly reduced in lung tissues in the model+DHA group(P<0.01,P<0.05,P<0.05).Whole gene transcriptome sequencing revealed that immune-related Plac8 and TLR7 genes were significantly upregu-lated(P<0.01)in mouse lung tissue of the model group but significantly downregulated(P<0.05)in mouse lung tissue of the model+DHA group.The results of RT-qPCR of Plac8 and TLR7 verified the results of whole gene transcriptome sequencing.GO and KEGG analysis showed that Plac8 and TLR7 were mainly related to the regulation of cytokine production,T/B cell activation and signal transduction,chemo-kine signal transduction and NF-κB signal transduction.CONCLUSION DHA might reduce LPS-induced lung damage and ameliorate the inflammatory condition in lungs of ALI mice.The mechanism of action may be that DHA negatively regulates the signaling pathways involved in TLR7 and Plac8 by decreasing the expressions of TLR7 and Plac8 mRNA before regulating a series of immune responses such as secretion of inflammation-related cytokines and activation of immune cells,thereby reducing inflam-matory damage in lungs.

Objective:To investigate the development of MLL-AF4 fusion gene-positive in newly-treated acute lymphoblastic leukemia (ALL) and to analyze the clinical characteristics and prognosis of patients with MLL-AF4 fusion gene-positive ALL.Methods:The clinical data of 455 patients with primary newly-treated ALL from February 2010 to February 2021 in Zhumadian Central Hospital were retrospectively analyzed. Fluorescence in situ hybridization was used to detect the presence of MLL gene rearrangement, and multiplexed nested reverse transcription polymerase chain reaction was used to detect the development of MLL-AF4 fusion gene-positive, and to compare the clinical characteristics and prognosis of patients with positive and negative MLL-AF4 fusion gene.Results:Among 455 patients with primary newly-treated ALL, 30 cases (6.6%) were positive for MLL gene rearrangement, including 21 positive for MLL-AF4 fusion gene (MLL-AF4 positive group), 6 cases positive for MLL-ENL fusion gene, 2 cases positive for MLL-AF9 fusion gene, and 1 case positive for MLL-AF10 fusion gene. A total of 60 ALL patients without any detection of fusion gene during the same period were randomly selected as the MLL-AF4-negative group. The proportion of those with hepatomegaly or splenomegaly, central nervous system (CNS) invasion, and the median peripheral blood leukocyte count and median peripheral blood na?ve cells absolute value in the MLL-AF4-positive group were higher than those in the MLL-AF4-negative group (all P < 0.05). The complete remission rates on day 15 of chemotherapy were 81.0% (17/21) and 90.0% (54/60), respectively in the MLL-AF4-positive group and MLL-AF4-negative group ( χ2 = 0.49, P = 0.484), and the cumulative relapse rates of 3 months were 38.1% (8/21) and 13.3% (8/60), respectively in the MLL-AF4-positive group and MLL-AF4-negative group ( χ2 = 4.56, P = 0.033). The overall survival in the MLL-AF4-positive group was worse than that in the MLL-AF4-negative group ( χ2 = 5.93, P = 0.015). Conclusions:Patients with MLL-AF4 fusion gene-positive ALL have a higher tumor load, a higher recurrence rate, a lower overall survival rate, and a poorer prognosis compared with negative patients at the initial diagnosis.

Objective:To explore the association between internet use and healthy lifestyles in urban adults.Methods:From May to August, 2022, a face-to-face questionnaire survey was conducted in residents aged 18-64 years selected in the urban area of Hangzhou by integrated cluster stratified random sampling and Kish grid method. The information about internet use included the internet use time in the past 7 days and 12 kinds of internet use contents. Using factor analysis and K-means clustering, three types of internet use were summarized, i.e. general type, video game type and working/learning type. Healthy lifestyles were defined as active physical activity, healthy diet habit, non-smoking, non-drinking, healthy weight, and healthy waist circumference. The correlations between internet use and healthy lifestyles were evaluated by using binary logistic regression and multinomial logistic regression analyses.Results:A total of 1 624 participants were included. After adjusting for potential confounding factors, the longer internet use time group (≥8.5 h/d) was less likely to have healthy weight ( OR=0.59, 95% CI:0.41-0.85) and 5-6 healthy lifestyles ( OR=0.55, 95% CI: 0.32-0.96) compared with those with shorter internet use time group (<2.5 h/d). For different types of internet use, it was found that compared with working/learning type group, the general type group was less likely to have healthy diet habits ( OR=0.63, 95% CI: 0.46-0.86), non-drinking ( OR=0.68, 95% CI: 0.47-0.99), healthy waist circumference ( OR=0.59, 95% CI: 0.42-0.84) and 5-6 healthy lifestyles ( OR=0.40, 95% CI: 0.23-0.69), the video game type group was less likely to have active physical activity ( OR=0.73, 95% CI: 0.55-0.97) and healthy diet habits ( OR=0.79, 95% CI: 0.62-0.99). Conclusion:Too long internet use (≥8.5 h/d), general type and video game type of internet use were associated with unhealthy lifestyles.

In 2022, World Health Organization (WHO) launched the 5th edition of the classification of renal tumors. This classification continues to use the previous classification framework of renal tumors based on morphology and tissue structure, and proposes the concept of renal cell carcinoma defined by molecular features for the first time. This article interprets from the 3 aspects of historical changes of WHO classification and grading of renal tumors, comparison of 2022 and 2016 WHO classification of renal tumors, and the role of molecular characteristics in the new pathological types such as ELOC mutant renal cell carcinoma, ALK rearrangement renal cell carcinoma, eosinophilic solid and cystic renal cell carcinoma. The purpose is to better understand the WHO from the traditional classification system based on tissue morphology to a three-in-one integrated classification system covering morphology, immunophenotype and genetic characteristics, and to understand the important value of molecular pathology in guiding the work of pathologists and clinicians under the new classification system.