Objective To investigate the effect and underlying mechanism of roxadustat on apoptosis and senescence of renal tubular epithelial cell line HK-2 induced by heat stress.Methods After HK-2 cells were treated with different concentrations of roxadustat(10,20,30,40 and 50 μmol/L)for 24 h,CCK-8 assay was used to determine the optimal intervention concentration of roxadustat.HK-2 cells were divided into 4 groups(n=3):control group,roxadustat group(30 μmol/L,24 h),heat-stress group(43 ℃,2 h),and heat-stress+roxadustat group(30 μmol/L roxadustat treatmnet for 24 h followed by heat-stress 2 h).Cell viability was detected by CCK-8 assay.Expression of hypoxia-inducible factor-1α(HIF-1α),Cleaved Caspase-3,p16 and p21 at protein level was detected by Western blotting.Immunofluorescence assay was employed to observe the distribution of HIF-1α.β-galactosidase staining kit was utilized to detect SA-β-Gal activity.TUNEL staining was used to measure cell apoptosis.Results The highest cell viability was observed in the cells after 30 μmol/L roxadustat treatment.Heat stress resulted in a significant decrease in cell viability(P＜0.05),elevated protein levels of HIF-1α,Cleaved Caspase-3,p16 and p21(P＜0.05),enhanced SA-β-Gal activity(P＜0.05)and increased percentage of TUNEL-positive cells(P＜0.05)when compared with the cells in the control group.In comparison with the heat-stress group,the heat-stress+roxadustat group showed significant decrease in the protein levels of Cleaved Caspase-3,p16 and p21(P＜0.05),reduced activity of SA-β-Gal[(65.44±5.00)％vs(77.15±2.61)％,P＜0.05]and decreased percentage of TUNEL-positive cells[(16.73±2.20)％vs(46.40±13.87)％,P＜0.05],but increase in cell viability[(86.33±4.51)％vs(66.33±8.50)％,P＜0.05]as well as HIF-1α protein expression(P＜0.05).Furthermore,immunofluorescence assay showed that HIF-1α was mainly distributed in the nucleus and perinucleus.Conclusion Roxadustat attenuates heat stress-induced apoptosis and senescence of renal tubular epithelial cells by upregulating HIF-1α.

Objective:To evaluate the influence of cytomegalovirus (CMV) infection on T cell senescence and cardiovascular disease (CVD) in maintenance hemodialysis (MHD) patients.Methods:It was a single center cross sectional study. Patients aged over 18 years old and received hemodialysis for at least 6 months at the Blood Purification Centre of the Department of Nephrology of Zhongshan Hospital Affiliated to Fudan University from January 2021 to April 2021 were enrolled. Demographic, hematological, nutritional and inflammatory markers were obtained. Anti-CMV-IgM and IgG antibodies were detected using the Roche Elecsys assay. CD28 - T cell was evaluated by flow cytometry. Mann-Whitney U test or Kruskal-Wallis H test was used for anti-CMV-IgG comparison among groups. Spearman correlation and linear regression were used to assess the relationship between anti-CMV-IgG and CD28 - T cell compartment. Logistic regression was used to assess the relationship between anti-CMV-IgG and CVD. Results:A total of 438 MHD patients (270 men and 168 women) were enrolled in the study. The median age was 62 (51, 70) years. The median time on hemodialysis was 57 (21, 100) months. The primary diseases included chronic glomerulonephritis [213 cases (48.6%)], diabetic nephropathy [82 cases (18.7%)], polycystic kidney disease [34 cases (7.8%)], hypertensive renal disease [34 cases (7.8%)], etc. Of these patients, 430 (98.2%) were seropositive for anti-CMV-IgG, 206 (47.0%) had anti-CMV-IgG titers exceeding the upper limit of 500 U/ml. Patients aged over 70 years old were 100% seropositive for anti-CMV-IgG. Patients on HD for more than 5 years had a higher seropositive rate of 99.1% than those with shorter HD duration, although these results were not statistically significant. Spearman correlation analysis showed that the anti-CMV-IgG titers in MHD patients were positively correlated with the proportion of CD4 + CD28 - T cells and CD8 + CD28 - T cells ( r=0.316, P<0.001; r=0.272, P<0.001). Multiple linear regression analysis showed that after adjusting for age and gender, lg[CD4 + CD28 - T cells(%)] and lg[CD8 + CD28 - T cells(%)] were positively correlated with lg[anti-CMV-IgG titers (U/ml)], respectively ( β=0.455, t=8.315, P<0.001; β=0.412, t=7.282, P<0.001). In analyzing the relationship between anti-CMV-IgG titers and CVD, patients were divided into six groups according to age and anti-CMV-IgG level. Group 1 included young patients with a lower anti-CMV-IgG titers (age ≤55 years old, anti-CMV-IgG <400 U/ml); Group 2 included young patients with a higher anti-CMV-IgG titers (age≤55 years old, anti-CMV-IgG ≥400 U/ml); Group 3 included middle-aged patients with a lower anti-CMV-IgG titers (5565 years old, anti-CMV-IgG<400 U/ml); Group 6 included old aged patients with a higher anti-CMV-IgG titers (age>65 years old, anti-CMV-IgG≥400 U/ml). The incidence of CVD was significantly different among the six groups ( χ2=18.780, P=0.002) and patients aged over 55 years old with higher anti-CMV-IgG level had a higher CVD incidence compared with group 1 ( P<0.05). Multivariate logistic regression analysis showed that increased age ( OR=1.020, 95% CI 1.002-1.038, P=0.033), male ( OR=1.855, 95% CI 1.161-2.965, P=0.010), history of diabetes ( OR=1.867, 95% CI 1.145-3.046, P=0.012), increased lg[NT-proBNP(μg/L)] ( OR=2.848, 95% CI 1.816-4.467, P<0.001) and lg[anti-CMV-IgG (U/ml)] ( OR=3.183, 95% CI 1.582- 6.405, P=0.001) were independent factors associated with CVD in MHD patients. Conclusions:CMV infection is extremely common in MHD patients. Increased anti-CMV-IgG is related to T cell senescence and CVD complications in MHD patients.

OBJECTIVE: To investigate how the National Health Commission of China (NHCC)-recommended Chinese medicines (CMs) modulate the major maladjustments of coronavirus disease 2019 (COVID-19), particularly the clinically observed complications and comorbidities. METHODS: By focusing on the potent targets in common with the conventional medicines, we investigated the mechanisms of 11 NHCC-recommended CMs in the modulation of the major COVID-19 pathophysiology (hyperinflammations, viral replication), complications (pain, headache) and comorbidities (hypertension, obesity, diabetes). The constituent herbs of these CMs and their chemical ingredients were from the Traditional Chinese Medicine Information Database. The experimentally-determined targets and the activity values of the chemical ingredients of these CMs were from the Natural Product Activity and Species Source Database. The approved and clinical trial drugs against these targets were searched from the Therapeutic Target Database and DrugBank Database. Pathways of the targets was obtained from Kyoto Encyclopedia of Genes and Genomes and additional literature search. RESULTS: Overall, 9 CMs modulated 6 targets discovered by the COVID-19 target discovery studies, 8 and 11 CMs modulated 8 and 6 targets of the approved or clinical trial drugs for the treatment of the major COVID-19 complications and comorbidities, respectively. CONCLUSION The coordinated actions of each NHCC-recommended CM against a few targets of the major COVID-19 pathophysiology, complications and comorbidities, partly have common mechanisms with the conventional medicines.
COVID-19/physiopathology* ; Comorbidity ; Drugs, Chinese Herbal/therapeutic use* ; Humans ; Medicine ; Medicine, Chinese Traditional ; SARS-CoV-2

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has become one major threat to human population health. The RNA-dependent RNA polymerase (RdRp) presents an ideal target of antivirals, whereas nucleoside analogs inhibitor is hindered by the proofreading activity of coronavirus. Herein, we report that corilagin (RAI-S-37) as a non-nucleoside inhibitor of SARS-CoV-2 RdRp, binds directly to RdRp, effectively inhibits the polymerase activity in both cell-free and cell-based assays, fully resists the proofreading activity and potently inhibits SARS-CoV-2 infection with a low 50% effective concentration (EC

Objective:To evaluate the effect of acetaminophen on sepsis-associated encephalopathy (SAE) in mice and the relationship with ferroptosis.Methods:A total of 160 clean-grade healthy adult male C57BL/6J mice, aged 6 weeks, weighing 22-24 g, were divided into 4 groups ( n=40 each) using a random number table method: sham operation group (Sham group), sham operation+ acetaminophen group (Sham+ APAP group), SAE group and SAE+ acetaminophen group (SAE+ APAP group). The model of SAE was established by cecal ligation and puncture in anesthetized mice.Acetaminophen 100 mg/kg was intraperitoneally injected at 1 h before the model was established in group Sham+ APAP and group SAE+ APAP.The postoperative 7-day survival rate was recorded.At 24 h after operation, brain tissues were taken for examination of the pathological changes of neurons in hippocampal CA1 region.At 24 h after establishment of the model, ultrastructure was observed with a transmission electron microscope, the contents of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) in hippocampus were determined by colorimetry, and the expression of glutathione peroxidase (GPX4), cystine/glutamate antiporter (xCT) and 4-hydroxy-2-nonenal (4-HNE) was determined by Western blot. Results:Compared with group Sham, the postoperative 7-day survival rate of mice was significantly decreased, contents of ROS and MDA in hippocampus were increased, GSH content in hippocampus was decreased and expression of GPX4 and xCT was down-regulated in SAE and SAE+ APAP groups, 4-HNE expression was up-regulated in group SAE ( P<0.05), and no significant change was found in the parameters mentioned above in group Sham+ APAP ( P>0.05). Compared with group SAE, the postoperative 7-day survival rate of mice was significantly increased, contents of ROS and MDA in hippocampus were decreased, GSH content in hippocampus was increased, expression of GPX4 and xCT was up-regulated and expression of 4-HNE was down-regulated in group SAE+ APAP ( P<0.05). The pathological changes in hippocampal CA1 region and damage to ultrastructure of hippocampal tissue were significantly attenuated in group SAE+ APAP as compared with group SAE. Conclusion:Acetaminophen can effectively alleviate SAE, and the mechanism is related to inhibiting ferroptosis in mice.

Objective To clarify the age patterns and types of differences so as to provide reference on prevention and interventions of hand,foot and mouth disease (HFMD) cases,in Hubei province.Methods We collected the HFMD case information of Hubei province from the Chinese National Notifiable Infectious Disease Reporting System in 2009-2015 while the information on pathogens from the laboratory monitoring system of Center for Disease Control and Prevention at all levels in Hubei province.All the data were stratified by age,disease severity,laboratory confirmation status,and serotypes of enterovirus.Results There were 495 783 reported HFMD cases from 2009 to 2015,in Hubei province,of which 1 045 were severe with 99 fatal.The annual notification rate was 1 231.0/106.HFMD cases were concentrated mainly in 0.5-5 year olds,with highest severity and mortality seen in 6-11 month-olds.The predominated pathogen in mild laboratory-confirmed cases each year,in order during 2009-2015 as:EV71,Cox A16,Cox A16,Cox A16,EV71,Cox A16 and other EV.HFMD showed semiannual peaks in April-June,November-December,and with more cases in the even years than in the odd years.Conclusions Children aged 0.5 to 5 years with 6 to 11 month-olds in particular,were the focused groups of attention in Hubei province.Our findings provided evidence for the improvement on monitoring program.Targeted intervention approaches should be strengthened to reduce the mortality and morbidity of HFMD in the province.

Objective To explore the risk factors of pulmonary artery hypertension (PAH) and the its relationship with T cell subsets to provide a foundation for the prevention and treatment of PAH.Methods 154 maintained hemodialysis (MHD) patients in our dialysis center were recruited according to the criterion and divided into two groups subsequently:PAH group (pulmonary artery systolic pressure,PASP ＞ 35 mmHg) and non-PAH group (PASP≤35 mmHg).The related clinical,biochemical and ultrasonic cardiogram data were collected and peripheral blood was acquired to detect the expressions of the surface antigen CD3,CD4,CD8 and CD69 with flow cytometry.Logistic regression analysis was used to find out the relationship between PAH and T cell subsets.Results There was no significant difference between 56 cases of PAH and 98 cases of non-PAH as regards gender,age,mean systolic and diastolic pressure,dialysis durations,morbidities of hypertension and diabetes,smoking rate,and left ventricular diameter.Compared with the non-PAH group,the PAH group demonstrated a lower percent of CD8 T cells and CD8 CD69 T cells,but a much higher left atrial diameter (LAD),Interventricular septum thickness,left ventricular posterior wall thickness,and NT-proBNP.The percentage of T cells,CD4 T cells and CD4 CD69 T cells showed no difference between the two groups.Multivariate analysis confirmed that PAH was negatively independently associated with the percentage of CD8 T cells and CD8CD69 T cells.Conclusions The decreased percentage of CD8 T cells and CD8CD69 T cells in the peripheral blood is a risk factor of PAH in maintained hemodialysis patients,and CD8 T cells may play an important role in the genesis of PAH.

Objective To investigate the relationship between the indexes change of lactate dehydrogenase(LDH),serum tumor associated material(BXTM),β2-microglobulin(β2-MG),C-reactive protein(CRP) and D-dimer(DD) with lymphoma cell bone marrow infiltration(BMI) degree in the patients with non-Hodgkin's lymphoma(NHL).Methods Seventy-four cases of initially treated NHL were retrospectively analyzed and divided into the bone marrow invasion group(BMI,26 cases),lymphoma cell leukemia group(LMCL,31 cases) and non-BMI group(N-BMI,17 cases).The related indexes levels in each group were detected.ROC curve was drawn and analyzed.Results The levels of LDH,β2-MG,CRP and DD levels in the BMI group and LMCL group were higher than those in the N-BMI group(P＜0.05).Based on the ROC curve analysis,the optimal diagnostic cut-off points of each index in BMI were determined.The CRP and LDH had good sensitivity(100.00 ％),specificity(80.00 ％-100.00 ％) and diagnostic accuracy(80.00 ％ 100.00％).In the LMCL group,the diagnostic accuracy of DD,BXTM and β2-MG was 63.30 ％-100.00％.Conclusion The changes of LDH,CRP,β2-MG and DD in the patients with NHL are consistent with the BMI severity of lymphoma cells,which has a good clinical value in staging,curative effect judgment and prognosis.

Objective:To investigate the expression of IL-6 and STAT3 in the epilepsy rat caused by penicillin .Methods:The model of epilepsy in rats was made by abdominal injecting penicillin .Then the EEG and behavior was observed in each group and the level of IL-6/STAT3 transcription and cytokine IL-6 secretion in different groups was compared using qRT-PCR and ELISA respectively.The protein expression of STAT3 was detected using Western blot.Results: 500 U/kg and 800 U/kg rats injected penicillin were induced epilepsy and show typical epileptic seizures behavioral and brain waves ,seizures was 100%;the level of IL-6/STAT3 transcription and IL-6 secretion in the model of epilepsy was significantly higher than control .There was no significant difference in the expression of STAT3 protein levels,but a significant increase in p-STAT3.Conclusion:Expression of STAT3 and IL-6 plays an important role in the Epilepsy Rat caused by penicillin .

Objective:To investigate the influence of water tank on endotoxin(ET)content in hemodialysis fluid.Methods:Specimens of hemodialysis fluid were collected from 36 hemodialysis centers in Shanghai from Nov 2010 to Oct 2012,and the ET content in hemodialysis fluid were detected by chromogenic substrate method.Results:A total of 2694 samples have been obtained from 36 hemodialysis centers during the 2 years’period.The contents of ET below 0.1 EU/mL were detected in 2348 (87.16%)samples,between 0.1 EU/mL and 0.5 EU/mL in 260(9.65%)samples,and above 0.5 EU/mL in 86(3.19%) samples.Water tanks were used in 19 of the 36 hemodialysis centers.ET contents of hemodialysis fluid in the hemodialysis cen-ters which used water tanks were higher than those didn’t use water tanks,and the difference showed statistical significance (P <0.05).Conclusion:The usage of water tank could significantly increase the ET content in hemodialysis fluid.Monitoring of water storage equipment should be strengthened.