BACKGROUND: Chronic kidney disease (CKD)-associated anemia (CKD-anemia) is associated with poor survival, and hemoglobin targets are often not achieved with current therapies. Phase 3 trials have demonstrated the treatment efficacy of roxadustat for CKD-anemia. This phase 4 study aims to evaluate the long-term (52-week) safety and effectiveness of roxadustat in a broad real-world patient population with CKD-anemia with and without dialysis in China. METHODS: This Phase 4 multicenter, open-label, prospective study, conducted from 24 November 2020 to 11 November 2022, evaluated the long-term safety and effectiveness of roxadustat for CKD-anemia in China. Patients aged ≥18 years with CKD-anemia with or without dialysis were included. The initial oral dose was 70-120 mg (weight-based followed by dose adjustment) over 52 weeks. The primary endpoint was safety based on adverse events (AEs). The secondary endpoints were hemoglobin changes from baseline and the proportion of patients who achieved mean hemoglobin ≥100 g/L. Effectiveness evaluable populations 1 (EE1) and EE2 included roxadustat-naïve and previously roxadustat-treated patients, respectively. The safety analysis set (SAF) included all patients who received ≥1 occasion. RESULTS: The EE1, EE2, and SAF populations included 1804, 193, and 2021 patients, respectively. In the SAF, the mean age was 50 ± 14 years, and 1087 patients (53.8%) were male. Mean baseline hemoglobin was 96.9 ± 14.0 g/L in EE1 and 100.3 ± 12.9 g/L in EE2. In EE1, the mean (95% confidence interval) hemoglobin changes from baseline over weeks 24-36 and 36-52 were 14.2 (13.5-14.9) g/L and 14.3 (13.5-15.0) g/L, respectively. Over weeks 24-36 and 36-52, 83.3% and 86.1% of patients in EE1 and 82.7% and 84.7% in EE2 achieved mean hemoglobin ≥100 g/L, respectively. In the SAF, 1643 (81.3%) patients experienced treatment-emergent AEs (TEAEs). Overall, 219 (10.8%) patients experienced drug-related TEAEs. Thirty-eight (1.9%) patients died of TEAEs (unrelated to the study drug). Vascular access thrombosis was uncommon. CONCLUSIONS: Roxadustat (52 weeks) increased hemoglobin and maintained the treatment target in Chinese patients with CKD-anemia with acceptable safety, supporting its use in real-world settings. REGISTRATION Chinese Clinical Trial Registry ( www.chictr.org.cn ) ChiCTR2100046322; CDE ( www.chinadrugtrials.org.cn ) CTR20201568.
Humans ; Male ; Female ; Anemia/etiology* ; Middle Aged ; Renal Insufficiency, Chronic/complications* ; Glycine/adverse effects* ; Isoquinolines/adverse effects* ; Aged ; Prospective Studies ; Adult ; Hemoglobins/metabolism* ; Treatment Outcome ; China ; Registries ; East Asian People

Objective To investigate the effect of programmed cell death-ligand 1(PD-L1)monoclonal antibody and paclitaxel(PTX)combined with lentinan(LNT)on human breast cancer MDA-MB-231 cells in vitro.Methods MDA-MB-231 cells,human peripheral blood mononuclear cells(PBMCs),and MDA-MB-231+PBMCs co-culture were randomly divided into control group,PTX group,LNT group,MPDL3280A(PD-L1 monoclonal antibody)group,PTX+LNT group,and PTX+LNT+MPDL3280A group.The cell activity was detected by CCK8.The expressions of MHC-I and PD-L1 were detected by flow cytometry.The levels of IFN-γ and TNF-α were detected by ELISA kits.Results Compared with control group,the activity of MDA-MB-231 cells was significant inhibited in PTX group,MPDL3280A group,PTX+LNT group,and PTX+LNT+MPDL3280A group(P＜0.01).LNT group and PTX+LNT+MPDL3280A group significantly promoted the immune effect of PBMCs(P＜0.05,P＜0.01).PTX+LNT+MPDL3280A group significantly inhibited the activity of MDA-MB-231cells co-cultured with PBMCs(0.56±0.16 vs.0.39±0.13,P＜0.05).Compared with the negative control,LNT significantly promoted the expression of PD-L1 in MDA-MB-231 and the secretion of IFN-γ by PBMCs(P＜0.05).Conclusion By blocking the effect between PD-L1 and PD-1,PD-L1 monoclonal antibody can improve immunity and promote the antitumor effect of PTX combined with LNT in vitro.

Objective To investigate the effect of programmed cell death-ligand 1(PD-L1)monoclonal antibody and paclitaxel(PTX)combined with lentinan(LNT)on human breast cancer MDA-MB-231 cells in vitro.Methods MDA-MB-231 cells,human peripheral blood mononuclear cells(PBMCs),and MDA-MB-231+PBMCs co-culture were randomly divided into control group,PTX group,LNT group,MPDL3280A(PD-L1 monoclonal antibody)group,PTX+LNT group,and PTX+LNT+MPDL3280A group.The cell activity was detected by CCK8.The expressions of MHC-I and PD-L1 were detected by flow cytometry.The levels of IFN-γ and TNF-α were detected by ELISA kits.Results Compared with control group,the activity of MDA-MB-231 cells was significant inhibited in PTX group,MPDL3280A group,PTX+LNT group,and PTX+LNT+MPDL3280A group(P＜0.01).LNT group and PTX+LNT+MPDL3280A group significantly promoted the immune effect of PBMCs(P＜0.05,P＜0.01).PTX+LNT+MPDL3280A group significantly inhibited the activity of MDA-MB-231cells co-cultured with PBMCs(0.56±0.16 vs.0.39±0.13,P＜0.05).Compared with the negative control,LNT significantly promoted the expression of PD-L1 in MDA-MB-231 and the secretion of IFN-γ by PBMCs(P＜0.05).Conclusion By blocking the effect between PD-L1 and PD-1,PD-L1 monoclonal antibody can improve immunity and promote the antitumor effect of PTX combined with LNT in vitro.

This study was designed to investigate the biological function and interaction mechanism of deubiquitinating enzyme USP14 and β-catenin in endometrial carcinoma(EC).The expression of USP14 and β-catenin in EC tissues and adjacent healthy tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry,while the effects of USP14 on β-catenin protein stability and ubiquitination were investigated by Western blotting and protein immunoprecipitation using HEA-1C and Ishikawa cell lines derived from EC.The effect of USP14 and β-catenin on EC cell proliferation was verified by cell proliferation assay(CCK-8);the regulatory effect of USP14 on β-catenin was verified by plasmid transfection.The regulation of USP14 and β-catenin on the expression of proteins related to epithelial-mesenchymal transition(EMT)in EC cells were verified by Western blotting.Data showed that the expression of USP14 and β-catenin in EC tissues were higher than those in adjacent healthy tissues,and USP14 and β-catenin interacted in EC cell line.USP14 maintained the protein stability of β-catenin through its deubiquitination activity.USP14-specific inhibitors could enhance the ubiquitination level of β-catenin and down-regulate the expression of β-catenin.USP14-specific inhibitors also inhibited EC cell proliferation and inhibited the expression of EMT-related proteins,while overexpression of β-catenin reversed the inhibition of EC cell growth and EMT induced by USP14 inhibitors.In conclusion,USP14 maintains β-catenin protein stability in EC cells by regulating the ubiquitination level,and the regulation of cell proliferation and EMT requires β-catenin.

This study was designed to investigate the biological function and interaction mechanism of deubiquitinating enzyme USP14 and β-catenin in endometrial carcinoma(EC).The expression of USP14 and β-catenin in EC tissues and adjacent healthy tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry,while the effects of USP14 on β-catenin protein stability and ubiquitination were investigated by Western blotting and protein immunoprecipitation using HEA-1C and Ishikawa cell lines derived from EC.The effect of USP14 and β-catenin on EC cell proliferation was verified by cell proliferation assay(CCK-8);the regulatory effect of USP14 on β-catenin was verified by plasmid transfection.The regulation of USP14 and β-catenin on the expression of proteins related to epithelial-mesenchymal transition(EMT)in EC cells were verified by Western blotting.Data showed that the expression of USP14 and β-catenin in EC tissues were higher than those in adjacent healthy tissues,and USP14 and β-catenin interacted in EC cell line.USP14 maintained the protein stability of β-catenin through its deubiquitination activity.USP14-specific inhibitors could enhance the ubiquitination level of β-catenin and down-regulate the expression of β-catenin.USP14-specific inhibitors also inhibited EC cell proliferation and inhibited the expression of EMT-related proteins,while overexpression of β-catenin reversed the inhibition of EC cell growth and EMT induced by USP14 inhibitors.In conclusion,USP14 maintains β-catenin protein stability in EC cells by regulating the ubiquitination level,and the regulation of cell proliferation and EMT requires β-catenin.

Objective To investigate the prevalence of soil-transmitted nematode human infections in Jurong City from 2016 to 2020, so as to provide the scientific evidence for formulating the control strategy. Methods During the period from 2016 to 2020, the permanent residents at ages of over 3 years living in Jurong City were selected as the study subjects. Stool samples were collected for the detection of soil-transmitted nematode eggs using the modified Kato-Katz thick smear method (two detections for one stool sample), and the species of hookworm was identified in stool-positive stool samples using the culture method. The prevalence and intensity of soil-transmitted nematode infections were calculated, and the change of the infection prevalence among years was examined using the Cochran-Armitage test for trend. Results A total of 10 011 people-time populations were detected for soil-transmitted nematode infections in Jurong City from 2016 to 2020, and 56 egg-positives were identified, with mean prevalence of 0.56%. The prevalence of soil-transmitted nematode human infections appeared a tendency towards a decline year by year in Jurong City (χ²_trend = 5.15, P < 0.01). The mean prevalence of hookworm, Ascaris lumbricoides and Trichuris trichiura infections was 0.44%, 0.11% and 0.20% in Jurong City from 2016 to 2020, respectively, and individuals with hookworm infections accounted for 78.57% of all cases with soil-transmitted nematode infections. Single parasite (98.21%) and mild infection were pre-dominant in individuals with soil-transmitted nematode infections, and no multiple infections were seen after 2016. Conclusions The prevalence of human soil-transmitted nematodiasis is low in Jurong City. Based on reinforcement of soil-transmitted nematodiasis surveillance, an increase in the health education investment is required to consolidate the control achievements.

【Objective】 To observe the uric acid-lowering effect of 3,4-dihydroxy-5-nitrobenzaldehyde (DHNB) on hyperuricemia models in mice and quails so as to improve the pharmacodynamic validation on hyperuricemia models. 【Methods】 The mouse hyperuricemia animal model was prepared by intraperitoneal injection of potassium oxonate 300 mg/kg; 30 g/(kg·d) yeast powder mixed feed (yeast powder∶feed, 1∶4) was used to prepare the quail hyperuricemia animal model. DHNB, 100 mg/kg, was intraperitoneally injected into the mice 1 hour prior to modeling; DHNB, 100 mg/kg, was intragastrically administered for two days consecutively into the quail hyperuricemia models. Control groups in mice and quails were set up respectively. Biochemical kits were used to detect serum uric acid, aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), creatinine (Cr), and blood urea nitrogen (BUN) in mouse and quail serum. Heart, lung, liver and kidney tissues of mice and quails were stained with HE. 【Results】 The serum uric acid in the mouse and quail hyperuricemia model groups was higher than that in the control group [(277.37±94.89) μmol/L vs. (176.49±44.83) μmol/L, P<0.05; (313.58±191.87) μmol/L vs. (167.26±66.56) μmol/ L, P<0.05)], indicating that the modeling was successful. DHNB could not reduce serum uric acid in hyperuricemia mouse model [(277.37±94.89) μmol/L vs. (238.72±63.43) μmol/L, P>0.05]. However, it significantly decreased serum uric acid in the quail model of hyperuricemia (313.58±191.87) μmol/L vs. (160.44±49.90)μmol/L, P<0.05]. Administration of DHNB 100 mg/kg one or two times had no effect on the liver and kidney functions of mice and quails, and had no toxicity to the heart, lung, liver or kidney tissues of mice and quails. 【Conclusion】 DHNB has a uric acid-lowering effect on the hyperuricemia quail model, and a single dose that caused the uric acid-lowering effect has no obvious toxicity to mouse or quail viscera. The quail hyperuricemia model is more suitable for the validation of the uric acid-lowering efficacy of DHNB than the mouse hyperuricemia model.

The aim of this paper was to study the specific mechanism of Fangji Huangqi Decoction(FHT) in decreasing uric acid and improving renal function in mice with hyperuricemia(HUA) induced by potassium oxonate, so as to provide theoretical basis for the research and development of drugs for clinical prevention and treatment of HUA and the modernization of traditional Chinese medicine. Sixty Kunming male mice were randomly divided into 6 groups, with 10 mice in each group, namely normal group, model group(250 mg·kg~(-1) potassium oxonate), FHT high, medium and low-dose groups(10 920, 5 460, and 2 730 mg·kg~(-1)) and positive drug allopurinol group(5 mg·kg~(-1)). Drug administration was given once a day for 7 days. On the 6 th day, mice of each group were kept in metabolic cages, and their urine was collected for 24 hours for determination of uric acid, creatinine, and β2-microglobulin(β2-MG) levels. After 7 days, the animals were sacrificed to determine serum uric acid, creatinine β2-MG and interleukin-1β(IL-1β) levels, and their liver and kidney tissues were collected. The liver tissues were used for subsequent determination of xanthine oxidase(XOD) activity, and the kidney tissues were used for subsequent determination of IL-1β levels, pathological tests and related Western blot experiments. In the cell transfection experiment, the cells were divided into blank group, model group(4.8 mmol·L~(-1) uric acid treatment), FHT administration group(4.8 mmol·L~(-1) uric acid+200 μg·mL~(-1) FHT), leucine-rich repeat kinase 1(LRRK1)-small interfering RNA(siRNA) group(4.8 mmol·L~(-1) uric acid+LRRK1-siRNA transfection) and LRRK1-siRNA+FHT group(4.8 mmol·L~(-1) uric acid+LRRK1-siRNA transfection+200 μg·mL~(-1) FHT). After 24 h incubation, the level of IL-1β in the cell supernatant was detected, and the cellular proteins were extracted and used to determine LRRK1, epidermal growth factor receptor(EGFR), PDZ kinase 1(PDZK1) and nuclear factor-kappa B(NF-κB) protein expression levels. The results showed that, FHT could significantly reduce the uric acid, creatinine and β2-MG levels in serum and β2-MG levels in urine, increase the uric acid and creatinine levels in urine, and improve the renal pathological results of the HUA mice, but showed no effect on liver XOD activity; at the same time, we found that the expression level of IL-1β in serum and kidney, NF-κB, LRRK1 and EGFR protein levels in kidney of HUA mice were significantly increased, and the expression level of PDZK1 protein was significantly decreased, while FHT could significantly improve the abnormal expression of these proteins, and FHT increased protein expression of renal organic anion transporter 1(OAT1), OAT3 and ATP bin-ding transporter G2(ABCG2) in HUA mice, but FHT had no effect on the expression of urate transporter 1(URAT1). In the cell transfection experiment, after transfection of LRRK1-siRNA, the levels of IL-1β, EGFR and NF-κB in supernatant were significantly reduced, and the expression of PDZK1 protein was significantly increased. As compared with the LRRK1-siRNA group, the levels of IL-1β, EGFR, PDZK1 and NF-κB did not change significantly with the additional FHT. This study showed that FHT may regulate the renal uric acid transport system through LRRK1 gene, improve the capacity of uric acid excretion, so as to reduce the level of serum uric acid. At the same time, FHT can not only protect the kidney directly, but also in an indirect manner by reducing the level of uric acid.
Animals ; Drugs, Chinese Herbal ; Hyperuricemia/drug therapy* ; Kidney ; Male ; Mice ; Uric Acid

Objective:To compare the changes of cerebral blood flow (CBF) in bipolar disorder type Ⅱ patients with and without suicidal ideation.Methods:Pseudo-continuous arterial spin labelling (pCASL) was examined with resting-state functional magnetic resonance imaging(rs-fMRI) on all subjects, CBF images of bipolar disorder type Ⅱ depression with suicidal ideation ( n=48), Bipolar Disorder type Ⅱ without suicidal ideation ( n=41), healthy controls ( n=62) at rest were collected. One-way ANOVA and Gaussian random field (GRF) were used to compare the CBF values between the groups to locate the regions with significant change. Results:Compared with the control group, CBF of left superior temporal gyrus (extending to the temporal pole) ( t=3.87, P<0.01; t=4.44, P<0.01) and left putamen ( t=4.04, P<0.01; t=3.19, P<0.01) increased in bipolar disorder type Ⅱ patients with and without suicidal ideation, and CBF of the right posterior cingulate gyrus (extending to the calcarine gyrus) decreased in bipolar disorder type Ⅱ patients with suicidal ideation( t=-4.66, P<0.01). Conclusions:There were abnormal CBF in left superior temporal gyrus and left putamen in bipolar disorder type Ⅱ patients with suicidal ideation and bipolar disorder type Ⅱ patients without suicidal ideation, while the decreased CBF in right posterior cingulate gyrus may be the specific brain change in bipolar disorder type Ⅱ patients with suicidal ideation.

Objective:To investigate the changes of white matter fiber bundles in patients with bipolar disorder depressive epoch.Methods:Forty-two patients with unmedicated bipolar disorder (BD) depression and 59 age-, sex- and handedness-matched healthy controls who underwent DTI were recruited in the study. According to the Johns Hopkins University human white matter fiber bundle map, the white matter tissue of the brain was segmented into 20 acknowledged large fiber bundles. The PANDA software was used to calculate the four average diffusion properties of each white matter fiber bundle for each subject. Nonparametric substitution test was used to detect the difference of diffusion index between the two groups on these 20 white matter fiber bundles. Pearson correlation analysis was performed between FA values and RD values extracted from significantly different white matter fiber bundles and clinical indices.Results:In comparison to the normal controls, BD patients had a significant decreased fractional anisotropy (FA) values in the left uncinate fasciculus (0.40±0.01 vs. 0.41±0.01, P=0.001) and the forceps minor (0.36±0.02 vs. 0.38±0.02, P<0.001). Additionally, the radial diffusivity values increased in the left uncinate fasciculus (6.57×10 -4±2.41×10 -5vs. 6.40×10 -4±2.42×10 -5, P=0.001 7). Pearson correlation analysis showed that there were no significant correlations among the clinical indices the FA values and AD values in the left uncinate fasciculus and forceps minor. Conclusions:The patients with bipolar disorder in depression possibly have abnormal left uncinate fasciculus and the forceps minor.