Objective:To discuss the effect of long non-coding RNA(lncRNA)DUXAP8 in exosomes(Exo)derived from the lung cancer A549 cells on the growth and immune escape of the lung cancer cells,and to clarify the mechanism.Methods:The human lung cancer cell line A549 was cultured,and its exosomes were extracted and identified.The A549 cells were treated with PKH67-labeled Exo to observe the uptake of Exo by A549 cells.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression level of lncRNA DUXAP8 in A549 cells before and after Exo treatment.The A549 cells were divided into control group(no treatment),Exo group(A549 cells treated with Exo),Exo+sh-NC group(A549 cells treated with Exo and then transfected with sh-NC),and Exo+sh-DUXAP8 group(A549 cells treated with Exo and then transfected with sh-DUXAP8).RT-qPCR method was used to detect the expression level of lncRNA DUXAP8 in A549 cells in various groups;colony formation assay was used to detect the colony formation abilities of the A549 cells in various groups;5-ethynyl-2'-deoxyuridine(EdU)staining method was used to detect the proliferation abilities of the A549 cells in various groups.After co-culturing A549 cells in various groups with human peripheral blood lymphocytes,flow cytometry was used to detect the percentages of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in various groups;3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)method was used to detect the killing rates of human peripheral blood lymphocytes on the A549 cells in various groups.Results:The diameter of Exo vesicles was 50-150 nm,and the exosome-specific marker proteins cluster of differentiation 63(CD63),cluster of differentiation 9(CD9),tumor susceptibility gene 101(TSG101),and heat shock protein 70(HSP70)were positively expressed,indicating successful exosome extraction.A549 cells efficiently took up PKH67-labeled Exo.The RT-PCR results showed that compared with A549 cells cultured alone,the expression level of lncRNA DUXAP8 in the A549 cells was increased after treatment with Exo derived from A549 cells(P<0.05).compared with control group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there were no significant difference in the expression level of IncRNA DUXAP8 in the cells in Exo+sh-NC group(P>0.05).The colony formation assay results showed that compared with control group,the number of colony formation of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the number of colony formation of the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the number of colony formation of the A549 cells in Exo+sh-NC group(P>0.05).The EdU staining results showed that compared with control group,the EdU-positive rate of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the EdU-positive rate in A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the EDU-positive rate in the cells in Exo+sh-NC group(P>0.05).The flow cytometry results showed that compared with control group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo group was decreased(P<0.05);compared with Exo group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo+sh-DUXAP8 group was increased(P<0.05),while there was no significant difference in the percentage of activated CD8+T lymphaytes in Exo+sh-NC group(P>0.05).The MTT assay results showed that compared with control group,the killing rate of human peripheral blood lymphocytes on the A549 cells in Exo group was decreased(P<0.05);compared with Exo group,the killing rate of human peripheral blood lymphocytes on A549 cells in Exo+sh-DUXAP8 group was increased(P<0.05),while no significant difference was observed in Exo+sh-NC group(P>0.05).Conclusion:The lncRNA DUXAP8 in exosomes derived from the lung cancer A549 cells promotes the proliferation of lung cancer cells and tumor immune escape.

Multi-rehabilitation treatment and early nutritional support are closely related to the recovery of critical ill patients.At present,a number of nutritional and rehabilitation interventions have been applied to critically ill patients to aid in their treatment and improve their nutritional status and motor dysfunction..In recent years,certain progress has been made in related research.This paper aims to summarize the latest literature on the application of multidisciplinary rehabilitation therapy in patients with CI under early nutritional support,provide comprehensive,systematic and practically valuable reference for clinical medical staff when formulating comprehensive treatment plans for CI patients under early nutritional support,and improve the treatment effect and quality of life of CI patients.

A method was developed for determination of dilauryl thiodipropionate(DLTDP)in fried foods by coupling solid-phase extraction(SPE)pretreatment with reverse-phase liquid chromatography-tandem mass spectrometry(RPLC-MS/MS)detection.Samples were extracted with n-hexane as the solvent,purified using a neutral alumina SPE cartridge,and finally analyzed by RPLC-MS/MS.Quantitative analysis was performed using matrix-matched calibration curves combined with an external standard method under optimal experimental conditions.The results showed that DLTDP exhibited good linearity in the range of 2.0-50.0 μg/L,with a correlation coefficient(R2)≥0.999.The limit of detection(LOD)and the limit of quantification(LOQ)of the method were 0.15 mg/kg and 0.5 mg/kg,respectively.The mean recoveries at three fortification levels(0.5,1.0,and 200 mg/kg)in different samples ranged from 84.8%to 96.8%,with the relative standard deviations(RSDs)all less than 8.0%.The developed method was highly sensitive,accurate and reliable,and easy to operate,making it well suited for the routine quantitative analysis of DLTDP in fried foods.

Objective:To observe the incidence of early blood eosinophils (Eos) elevation in patients with peritoneal dialysis (PD), analyze its related factors, and its relationship with early-onset peritonitis in PD patients.Methods:This study was a retrospective observational cohort study. Patients who underwent PD catheterization in Peking University People's Hospital from January 2012 to December 2022 were included. After surgery, PD treatment was started immediately and followed up regularly ≥12 months. The general information and laboratory indexes collected 1 week before catheterization, and at 1, 3, 6 and 12 months after catheterization were recorded. The occurrence of elevated blood Eos (≥0.5×10 9/L) during the early stage of PD, the related factors of Eos elevation (≥0.5×10 9/L) and the relationship with early-onset peritonitis (within 12 months after PD initiation) were analyzed. Results:(1) A total of 235 patients were enrolled, with an age of (57.9.±13.9) years, including 136 males (57.9%). The primary diseases were predominantly chronic glomerulonephritis (111/235, 42.7%) and diabetic nephropathy (83/235, 35.3%). During the 12-month follow-up period, 73 patients had elevated blood Eos (31.1%), of which 37 cases (50.7%) occurred within 1 month after PD catheterization, 21 cases (28.7%) occurred 2-3 months after PD catheterization, 12 cases (16.4%) occurred 4-6 months after PD catheterization, and 3 cases (4.1%) occurred 7-12 months after PD catheterization. In 73 patients with elevated Eos, 69 cases (94.5%) were mildly elevated, 4 cases (5.5%) were moderately elevated. As for the duration of elevated blood Eos, 28 cases (38.4%) lasted less than 1 month, 27 cases (37.0%) lasted 1-3 months, and 18 cases (24.7%) lasted more than 3 months. (2) In patients with elevated blood Eos, the proportion of male patients (71.4% vs. 52.1%, χ 2=7.515, P=0.006), the proportion of diabetes mellitus (55.7% vs. 41.2%, χ 2=4.168, P=0.046), and the proportion of combined vascular disease (32.9% vs. 18.2%, χ 2=6.060, P=0.017) were significantly higher than those patients in normal blood Eos group. (3) Multivariate Logistic regression analysis showed that male was an independent related factor for elevated blood Eos (≥0.5×10 9/L) in the early stages of PD ( OR=2.044, 95% CI 1.101- 3.794, P=0.023). (4) Diabetes mellitus ( OR=3.363, 95% CI 1.087-10.405, P=0.035), lower baseline hemoglobin level ( OR=0.941, 95% CI 0.903-0.980, P=0.004) and elevated blood Eos (with serum Eos<0.5×10 9/L as reference, OR=2.917, 95% CI 1.022-8.326, P=0.045) were the independent related factors of early-onset peritonitis. Conclusion:Blood Eos elevations are common in early stage of PD patients , mainly occuring within 6 months after PD catheterization, and most of them are slightly increased and last less than 3 months. Male sex is an independent related factor for the elevation of blood Eos in the early stage of PD. Elevated blood Eos is an independent related factor for early-onset peritonitis.

Objective:To observe the incidence of early blood eosinophils (Eos) elevation in patients with peritoneal dialysis (PD), analyze its related factors, and its relationship with early-onset peritonitis in PD patients.Methods:This study was a retrospective observational cohort study. Patients who underwent PD catheterization in Peking University People's Hospital from January 2012 to December 2022 were included. After surgery, PD treatment was started immediately and followed up regularly ≥12 months. The general information and laboratory indexes collected 1 week before catheterization, and at 1, 3, 6 and 12 months after catheterization were recorded. The occurrence of elevated blood Eos (≥0.5×10 9/L) during the early stage of PD, the related factors of Eos elevation (≥0.5×10 9/L) and the relationship with early-onset peritonitis (within 12 months after PD initiation) were analyzed. Results:(1) A total of 235 patients were enrolled, with an age of (57.9.±13.9) years, including 136 males (57.9%). The primary diseases were predominantly chronic glomerulonephritis (111/235, 42.7%) and diabetic nephropathy (83/235, 35.3%). During the 12-month follow-up period, 73 patients had elevated blood Eos (31.1%), of which 37 cases (50.7%) occurred within 1 month after PD catheterization, 21 cases (28.7%) occurred 2-3 months after PD catheterization, 12 cases (16.4%) occurred 4-6 months after PD catheterization, and 3 cases (4.1%) occurred 7-12 months after PD catheterization. In 73 patients with elevated Eos, 69 cases (94.5%) were mildly elevated, 4 cases (5.5%) were moderately elevated. As for the duration of elevated blood Eos, 28 cases (38.4%) lasted less than 1 month, 27 cases (37.0%) lasted 1-3 months, and 18 cases (24.7%) lasted more than 3 months. (2) In patients with elevated blood Eos, the proportion of male patients (71.4% vs. 52.1%, χ 2=7.515, P=0.006), the proportion of diabetes mellitus (55.7% vs. 41.2%, χ 2=4.168, P=0.046), and the proportion of combined vascular disease (32.9% vs. 18.2%, χ 2=6.060, P=0.017) were significantly higher than those patients in normal blood Eos group. (3) Multivariate Logistic regression analysis showed that male was an independent related factor for elevated blood Eos (≥0.5×10 9/L) in the early stages of PD ( OR=2.044, 95% CI 1.101- 3.794, P=0.023). (4) Diabetes mellitus ( OR=3.363, 95% CI 1.087-10.405, P=0.035), lower baseline hemoglobin level ( OR=0.941, 95% CI 0.903-0.980, P=0.004) and elevated blood Eos (with serum Eos<0.5×10 9/L as reference, OR=2.917, 95% CI 1.022-8.326, P=0.045) were the independent related factors of early-onset peritonitis. Conclusion:Blood Eos elevations are common in early stage of PD patients , mainly occuring within 6 months after PD catheterization, and most of them are slightly increased and last less than 3 months. Male sex is an independent related factor for the elevation of blood Eos in the early stage of PD. Elevated blood Eos is an independent related factor for early-onset peritonitis.

Objective:Peritoneal dialysis(PD)-associated peritonitis is a common and major complica-tion of PD and the most common cause of technical failure of PD.The presence of bacterial biofilm may be an important factor leading to refractory or recurrence of peritonitis.To investigate the formation and characteristics of bacterial biofilms on PD catheters after peritonitis-associated catheter removal.Methods:The patients with maintenance PD who were regularly followed up in the Peking University People's Hospital from June 2007 to January 2022 were retrospectively analyzed.The patients who with-drew from PD because of peritonitis and removed the PD catheter in our hospital and underwent the scan-ning electron microscope examination of the catheter were selected.The general information of the pa-tients,the electron microscope results of the PD catheter and the bacterial culture results of the PD fluid were summarized.Results:(1)A total of 18 patients were included,11 were female(accounting for 61.1％).The average age of the patients was(59.1±11.5)years,and the average duration of dialysis was(80.1±47.4)months.Primary kidney diseases were predominantly chronic glomerulonephritis(55.6％),followed by diabetic nephropathy(27.8％),and others(16.6％).The reasons for cathe-ters removal in 18 patients were refractory peritonitis in 11 cases,recurrent peritonitis in 5 cases,and fungal peritonitis in 2 cases.(2)16 of the 18 patients(88.9％)had catheter bacterial biofilm,and the bacterial biofilm forms were all cocci.Some were arranged in grape-like shapes,and their diameters ranged from about 500 nm to 1 000 nm.The bacterial culture results of peritoneal dialysis fluid showed that the three most common pathogens were Escherichia coli,methicillin-sensitive Staphylococcus aureus(MSSA),and Staphylococcus epidermidis.(3)Among the 18 patients enrolled,13 patients(72.2％)had peritonitis in the past.The causative bacteria of peritonitis in 9 patients were cocci,including coagu-lase-negative Staphylococci(Staphylococcus suis,Staphylococcus surface,Staphylococcus xylosus,Staphy-lococcus warneri),Staphylococcus aureus,Streptococcus(Streptococcus salivarius and Aerococus viridans).Conclusion:Bacterial biofilm formation on the inner surface of PD catheter is common in peritonitis-asso-ciated catheter removal patients.Not all PD catheters removed due to peritonitis have bacterial biofilms.Bacterial biofilms and peritonitis pathogens may not be consistent.

AIM To study endophytic fungi from Scutellaria baicalensis Georgi.and the enzyme inhibitory activities of their secondary metabolites.METHODS Six different media were used to isolate and purify endophytic fungi from S.baicalensis by tissue homogenate method.The activities of secondary metabolites were evaluated by targeting different enzymes.The highly active strains were identified by molecular biology combined with morphology,and the highly active chemical components were tracked and separated by modern chromatographic separation technology.RESULTS Sixty-four endophytic fungal strains were isolated from S.baicalensis,and one hundred and twenty-eight secondary metabolites were obtained by fermentation.The samples with certain inhibitory activities against adenosine deaminase(ADA),β-lactamase and tyrosinase(TYR)accounted for 14.06%,3.91%and 18.75%,respectively.Strain HTS-23-2 showed high TYR inhibitory activity,and 99%homology with Aspergillus flavus by molecular identification.One compound was isolated from the fermentation samples and identified as kojic acid.CONCLUSION S.baicalensis harbors a rich diversity of endophytic fungi,which serve as a valuable resource for active substances.

Objective:To investigate the changes of exam scores of retrained peritoneal dialysis (PD) operators (patients, family members, or nannies) with an internal of one year during COVID-19 epidemic and provide basis for targeted training.Methods:It was a cross-sectional survey study. The maintenance PD patients who participated in two trainings with an interval of one year during COVID-19 epidemic from November 1, 2019 to February 28, 2021 in Department of Nephrology in Peking University People's Hospital were enrolled. During COVID-19 epidemic, retraining was extended from once every six months to once a year. The clinical data were collected, the self-designed training exam score table including theoretical knowledge and operational skills assessment was used to investigate the exam scores of two trainings, and the total exam scores and sub-item scores of PD operators before and after one year were compared. Logistic regression analysis was used to analyze the associated factors of the reduction of exam scores.Results:A total of 59 patients were enrolled, with 35 males (59.32%), age of (58.41±14.52) years, and dialysis duration of 42 (12, 84) months. There were 54 patients (91.53%) operating by themselves, 22 operators (37.29%) having college degree or above, and 35 operators (59.32%) having decreased exam scores. The total exam scores were 83.17±7.90 and 80.61±8.20 before and after one year, respectively ( t=2.732, P=0.008). In the six contents of itemized scoring, compared with one year ago, the exam scores of complication treatment ( t=4.928, P<0.001) and self-monitoring ( t=3.222, P=0.002) were significantly decreased. There was no statistically significant difference in the exam scores of environment and hygiene, dialysate replacement operation, exit nursing and diet before and after one year (all P>0.05). The total exam scores in patients with dialysis duration <12 months and 36-60 months after one year were significantly lower than before one year ( t=2.309, P=0.041; t=3.086, P=0.009). There was no statistically significant difference in the exam scores of PD operators with dialysis duration of 12-<36 months and >60 months before and after one year (both P>0.05). Logistic regression analysis showed that dialysis duration was an independent associated factor of exam scores reduction (dialysis duration 36-60 months/>60 months, OR=6.233, 95% CI 1.035-37.529, P=0.046). Conclusions:During COVID-19 epidemic, the reduced frequency of retraining reduces the training exam scores of PD operators, especially in patients with dialysis duration of 36-60 months. The weak points are focused on complication management and self-monitoring. Training should be strengthened for key patients and key contents if regular retraining is not possible due to special circumstances.

Aim To establish a quantitative immunofluorescent bioactivity assay(ICW)for insulin glargine based on CHO-IN-SRB 1284 transgenic cells,and to study its equivalence with in-sulin bioassay of Ch.P.Methods The cells were diluted 25 times with 1.5 × 108 L-1 cell density plates and 1 500 μmol·L-1 insulin glargine,and then diluted with a 3-fold gradient se-ries.The cells were stimulated in microporous plates for 20 min.After fixation,permeation and antibody incubation.Quantitative immunofluorescence biological activity was detected by odyssey two-color infrared fluorescence imaging system.Results There was a good dose-effect relationship between the concentration of insulin glargine in ICW and its relative potency.The method had good specificity,and the relative accuracy,intermediate preci-sion and linearity met the requirements.The relative deviation of biological activity results of 7 batches of insulin glargine samples measured by the two methods was less than 10％.The results were analyzed by SPSS and SAS software,which showed that the methods were correlated and equivalent.Conclusions The quantitative immunofluorescence assay for the biological activity of insulin glargine can be established.The method has good spe-cificity,high accuracy and precision,and has correlation and e-quivalent with biotiter assay,which can be applied to in vitro ef-ficacy evaluation and quality control of insulin glargine.

Objective To observe the inter-observer consistency of diameter of inferior vena cava(IVC)and IVC collapsibility index(IVCCI)measured and assessed with echocardiography and the correlations with right heart parameters in patients with right heart dysfunction.Methods Forty-seven patients with right heart dysfunction were prospectively recruited in observation group,while 50 adults with normal right heart function were taken as controls(control group).Parameters of the right heart were obtained with echocardiography,including the right ventricular fractional area change(FAC),the tricuspid annular plane systolic excursion(TAPSE),the myocardial performance index(MPI),the tricuspid annular systolic velocity(S')as well as early and late diastolic velocity(e',a')and e'/a' ratio,also the tricuspid valve orifice early and late diastolic velocities(E,A)and E/A ratio and E/e',the vena contracta width of tricuspid regurgitation(TR-VCW),the maximum velocity of tricuspid regurgitation(TR-Vmax),the pulmonary artery systolic pressure(PASP)and right atrial area(RAA).Besides,the maximal and minimal diameter of IVC(IVCDmax,IVCDmin)during the respiratory cycle were measured with two dimensional(2D)ultrasound and anatomical M-mode ultrasound,respectively,and the IVCCI were calculated.Then 20 subjects were randomly selected from each group,and IVC parameters were obtained.The basic data,right heart parameters and IVC parameters were compared between groups,intra-class correlation coefficient(ICC)between 2 sonographers of IVC parameters were calculated,and correlations between IVC parameters and right heart parameters were assessed.Results No significant differences of gender,age nor body mass index(BMI)was detected between groups(all P＞0.05).Compared with those in control group,MPI,e',e'/a',E,A,E/e',TR-VCW,TR-Vmax,PASP and RAA increased,whereas FAC,TAPSE,S'and a'decreased in observation group(all P＜0.05).The inter-observer consistencies were good for IVCDmax and IVCCI in observation group(ICC=0.787-0.971)and IVCDmax in the control group(ICC=0.971,0.964)obtained with 2D ultrasound and anatomical M-mode ultrasound,but poor for IVCCI in control group(ICC=0.169,0.456).Compared with those in control group,IVC parameters 2D-IVCDmax,2D-IVCDmin,M-IVCDmax and M-IVCDmin increased but 2D-IVCCI and M-IVCCI decreased in observation group(all P＜0.05).In control group,2D-IVCDmax was weakly negatively correlated with TAPSE and a'(r=-0.392,-0.364),weakly positively correlated with e'/a',E,E/A,TR-VCW and RAA(r=0.396,0.483,0.461,0.565,0.582),2D-IVCCI was weakly negatively correlated with TR-VCW and RAA(r=-0.386,-0.380),while M-IVCDmax was weakly negatively correlated with TAPSE(r=-0.384),and weakly positively correlated with e'/a',E,E/A,TR-VCW and RAA(r=0.357,0.453,0.473,0.549,0.550),M-IVCCI was weakly negatively correlated with MPI,E,TR-VCW and RAA(r=-0.347,-0.337,-0.475,-0.421).Conclusion In patients with right heart dysfunction,IVCD diameter and IVCCI obtained with echocardiography had good inter-observer consistencies.Parameters obtained with 2D ultrasound and anatomic M-mode ultrasound had certain relations with the right heart parameters.