To investigate the role of intestinal fungi in the progression of heart failure (HF) associated with chronic kidney disease (CKD).

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 programmed death including necroptosis,apoptosis,autophagy,ferroptosis,and pyroptosis in bone marrow megakaryocytes of mice during sepsis and its impact on platelet production capacity and coagulation function in mice.Methods C57BL/6J mice were randomly divided into a sham operation group(sham group)and a sepsis model group(CLP group).Peripheral blood platelets and coagulation function were measured by abdominal aortic blood sampling at 24 h postoperatively in both sham and CLP groups.After the mice were sacrificed,long bones of both lower limbs were taken,and bone marrow megakaryocytes were extracted using megakaryocyte separation solution and immunomagnetic bead separation.Laser confocal microscopy was used to observe the activation of programmed death-related marker molecules in mouse bone marrow megakaryocytes.Flow cytometry was used to detect programmed death rate,platelet production phenotype,and platelet surface markers(CD41,CD42b,CD61)of megakaryocytes.Western blotting was used to detect the expression of programmed death-related proteins in megakaryocytes.Results Compared with the sham group,the CLP group showed significant decreases in the number of platelets during acute sepsis(24 h)(P<0.000 1),significant increases in platelet distri-bution width(PDW)and mean platelet volume(MPV)(P<0.01),significant prolonging of thrombin time(TT),prothrombin time(PT),and activated partial thromboplastin time(APTT)(P<0.000 1,P<0.001,P<0.01),and significant reduction in fibrinogen(Fib)(P<0.000 1).Compared with the Con/sham group,the LPS/CLP group exhibited significant increases in the platelet production phenotype of megakaryocyte,the number of PLP in the supernatant,and the expression levels of platelet surface markers(CD41,CD42b,CD61).The rates of megakaryocyte necroptosis/apoptosis,pyroptosis,and ferroptosis were significantly elevated at 24 h post-CLP surgery.Laser confo-cal microscopy showed significant activation of LC3,P-MLKL,Caspase-1,and Fe2+in megakaryocytes of mice after CLP surgery.Western blotting results revealed that the CLP group exhibited a significant increase in the activa-tion rate of necroptosis-related protein P-MLKL(P<0.001),a significant increase in the cleavage of pyroptosis-related proteins GSDMD and GSDMD-N(P<0.01,P<0.001,respectively),a significant increase in the expres-sion of ferroptosis-related protein ACSL4(P<0.01),and a significant decrease in the expression of GPX4(P<0.01)compared to the sham group.Additionally,the CLP group demonstrated significant increases in the expression of apoptosis-related protein Bax,the cleavage of autophagy-related protein LC3B-Ⅱ,and the expression of P62(P<0.05,P<0.001,P<0.001,respectively).Inhibition of apoptosis with programmed cell death inhibitors decreased platelet production function of megakaryocyte,while inhibition of necroptosis and pyroptosis had limited effects on platelet production function of megakaryocyte.Inhibition of ferroptosis and autophagy enhanced platelet production function of megakaryocyte.Conclusion Significant programmed death of megakaryocytes was observed during the acute phase of sepsis(24 h).Among those megakaryocytes,apoptosis is an important mechanism for the differentia-tion of platelet production phenotype and increased platelet production capacity of megakaryocyte.Overactive autophagy and ferroptosis in megakaryocytes lead to megakaryocyte dysfunction,which is an important mechanism for coagulation abnormalities in sepsis.

Objective To investigate programmed death including necroptosis,apoptosis,autophagy,ferroptosis,and pyroptosis in bone marrow megakaryocytes of mice during sepsis and its impact on platelet production capacity and coagulation function in mice.Methods C57BL/6J mice were randomly divided into a sham operation group(sham group)and a sepsis model group(CLP group).Peripheral blood platelets and coagulation function were measured by abdominal aortic blood sampling at 24 h postoperatively in both sham and CLP groups.After the mice were sacrificed,long bones of both lower limbs were taken,and bone marrow megakaryocytes were extracted using megakaryocyte separation solution and immunomagnetic bead separation.Laser confocal microscopy was used to observe the activation of programmed death-related marker molecules in mouse bone marrow megakaryocytes.Flow cytometry was used to detect programmed death rate,platelet production phenotype,and platelet surface markers(CD41,CD42b,CD61)of megakaryocytes.Western blotting was used to detect the expression of programmed death-related proteins in megakaryocytes.Results Compared with the sham group,the CLP group showed significant decreases in the number of platelets during acute sepsis(24 h)(P<0.000 1),significant increases in platelet distri-bution width(PDW)and mean platelet volume(MPV)(P<0.01),significant prolonging of thrombin time(TT),prothrombin time(PT),and activated partial thromboplastin time(APTT)(P<0.000 1,P<0.001,P<0.01),and significant reduction in fibrinogen(Fib)(P<0.000 1).Compared with the Con/sham group,the LPS/CLP group exhibited significant increases in the platelet production phenotype of megakaryocyte,the number of PLP in the supernatant,and the expression levels of platelet surface markers(CD41,CD42b,CD61).The rates of megakaryocyte necroptosis/apoptosis,pyroptosis,and ferroptosis were significantly elevated at 24 h post-CLP surgery.Laser confo-cal microscopy showed significant activation of LC3,P-MLKL,Caspase-1,and Fe2+in megakaryocytes of mice after CLP surgery.Western blotting results revealed that the CLP group exhibited a significant increase in the activa-tion rate of necroptosis-related protein P-MLKL(P<0.001),a significant increase in the cleavage of pyroptosis-related proteins GSDMD and GSDMD-N(P<0.01,P<0.001,respectively),a significant increase in the expres-sion of ferroptosis-related protein ACSL4(P<0.01),and a significant decrease in the expression of GPX4(P<0.01)compared to the sham group.Additionally,the CLP group demonstrated significant increases in the expression of apoptosis-related protein Bax,the cleavage of autophagy-related protein LC3B-Ⅱ,and the expression of P62(P<0.05,P<0.001,P<0.001,respectively).Inhibition of apoptosis with programmed cell death inhibitors decreased platelet production function of megakaryocyte,while inhibition of necroptosis and pyroptosis had limited effects on platelet production function of megakaryocyte.Inhibition of ferroptosis and autophagy enhanced platelet production function of megakaryocyte.Conclusion Significant programmed death of megakaryocytes was observed during the acute phase of sepsis(24 h).Among those megakaryocytes,apoptosis is an important mechanism for the differentia-tion of platelet production phenotype and increased platelet production capacity of megakaryocyte.Overactive autophagy and ferroptosis in megakaryocytes lead to megakaryocyte dysfunction,which is an important mechanism for coagulation abnormalities in sepsis.

Medicinal plants are a valuable source of essential medicines and herbal products for healthcare and disease therapy. Compared with chemical synthesis and extraction, the biosynthesis of natural products is a very promising alternative for the successful conservation of medicinal plants, and its rapid development will greatly facilitate the conservation and sustainable utilization of medicinal plants. Here, we summarize the advances in strategies and methods concerning the biosynthesis and production of natural products of medicinal plants. The strategies and methods mainly include genetic engineering, plant cell culture engineering, metabolic engineering, and synthetic biology based on multiple "OMICS" technologies, with paradigms for the biosynthesis of terpenoids and alkaloids. We also highlight the biosynthetic approaches and discuss progress in the production of some valuable natural products, exemplifying compounds such as vindoline (alkaloid), artemisinin and paclitaxel (terpenoids), to illustrate the power of biotechnology in medicinal plants.

Objective To evaluate the quality of ultrafine powder of Spatholobus suberectus from different sources by combined HPLC fingerprint and chemical pattern recognition.Methods Separation was performed on a Amethyst C18-H column(4.6 mmx250 mm,5 pm)and the mobile phase was 0.1％phosphoric acid-acetonitrile with gradient elution.The flow rate was 1.0 mL/min and the wavelength was 260 nm.The column temperature was 30 ℃.The cluster analysis(CA),least partial squares-discriminant analysis(PLS-DA)and principal component analysis(PCA)were applied to studying the HPLC fingerprint and chemical pattern recognition.Results Thirteen common peaks were identified in ultrafine powder samples of 10 batches of Spatholobus suberectus and 3 components were identified by reference substances.The similarity of fingerprint was 0.921-0.989,indicating good similarity.The samples were divided into 3 groups by cluster analysis;principal component analysis results extracted 4 principal components.According to the composite score,the quality of ultrafine powder samples of Spatholobus suberectus from S6,S1,S2,S3 were better than others.And 4 kinds of marker compounds that caused the quality difference of ultrafine powder samples of Spatholobus suberectu were screened out through the least partial squares-discriminant analysis,which were epicatechin and peaks 1,6 and 4 respectively.Conclusion The fingerprint method is simple and reproducible,and can be used to evaluate the quality of ultrafine powder samples of Spatholobus suberectus in combination with multi-mode stoichiometric analysis.

Objective:To explore the application effect of key node advanced nursing mode in the treatment of patients with acute myocardial infarction.Methods:In October 2020, the hospital established a Critical Control Point rescue mode management team.122 patients with acute myocardial infarction admitted to emergency department of the hospital were enrolled as the objects between October 2020 and October 2021. The healthcare failure mode and effect analysis model was applied to analyze the shortcomings of emergency process, so as to construct critical control point rescue mode in the treatment of patients with acute myocardial infarction and apply it to the clinic in November 2021. After clinical application, emergency nursing and cardiac function recovery were compared between the two groups. The mortality rate within 30 d after surgery and occurrence of complications during hospitalization were recorded.Results:The first medical contact to balloon time dropped from (81.9±6.54) min to (56.2±4.23)min. The time from first medical contact to diagnosis of acute myocardial infarction dropped from (47.3±5.68) min to (30.69±5.21) min, the door-balloon dilation time dropped from (49.79±13.84) min to (28.63±15.71) min, producing results time of myocardial injury markers dropped from (28.38±3.79)min to (19.26±2.17) min, reporting time of electrocardiogram dropped from (5.82±2.01) min to (5.14±1.89)min, and hospitalization time dropped from (7.25±2.18) min to (6.14±1.27) min, and the differences were statistically significant ( P<0.05). After treatment, left ventricular ejection fraction in observation group was higher than that in control group, left ventricular end-diastolic diameter and cardiac troponin were lower than those in control group ( P<0.05). The incidence of hypotension and malignant arrhythmia in observation group was lower than that in control group ( P<0.05). Conclusions:The critical control point rescue mode can shorten treatment time and hospitalization time in acute myocardial infarction patients, improve cardiac function, and reduce the risk of complications during hospitalization.

Objective To investigate the causes of abnormal decrease in maximum amplitude(MA)of thromboelastog-raphy(TEG)and its effect on prognosis by monitoring the changes of coagulation-related indexes in emergency trauma pa-tients.Methods A total of 319 cases of trauma patients admitted to our hospital from September 2020 to September 2023 were retrospectively analyzed,and the coagulation-related indexes of 0 h and 24 h after admission were observed.According to the MA results,they were divided into normal MA group(>50 mm)and reduced MA group(≤50 mm)to compare the hemoglobin(Hb),platelets count(Plt),activated partial thromboplastin time(APTT),prothrombin time(PT),fibrinogen(Fib),thrombin time(TT),D-dimer(D-D),coagulation reaction time(R),clot formation kinetics(Angle),30 min clot dissolution rate(Ly30),MA,thrombine-antithrombin complex(TAT)and plasminase-α2 plasminase inhibitor complex(PIC).The correlation between MA and fibrinolysis indexes in 319 trauma patients was analyzed.According to whether tranexamic acid(TXA)was used,the reduced MA group was divided into a TXA group and a non-drug group.The differ-ences in the change of the above coagulation-related indexes,mortality rate and changes in blood product dosage were com-pared between the two groups.Results Compared with the normal MA group,Hb,Plt,Fib,diastolic blood pressure and GCS scores decreased,while heart rate,ISS score and mortality increased significantly in the reduced MA group(P<0.05).The R,PT and TT were prolonged significantly(P<0.05),and PIC and D-D increased significantly(P<0.05)in the re-duced MA group.Correlation analysis found that MA had no correlation with Ly30,TAT and APTT,but was correlated with Angle(r=0.803),Plt(r=0.544),Fib(r=0.581),PIC(r=-0.443)and D-D(r=-0.343).Compared with the non-drug group,the change of Angle,MA and FIB in the TXA group increased significantly(P<0.05),while the change of PIC de-creased(P<0.05).Cryoprecipitate and platelet transfusion in the TXA group reduced significantly(P<0.05),and red blood cell transfusion had a decreasing trend,but the difference was not significant(P>0.05).The mortality rate in the TXA group was reduced significantly(P<0.05).Conclusion Hyperfibrinolysis may be an important factor in the abnormal decrease of MA in emergency trauma patients.Treatment with TXA can improve its effect on MA,and reduce the transfusion of blood products and the patient mortality.

Objective To investigate preliminary clinical effects of three-dimensional(3D)printing combined with minimally invasive percutaneous plate osteosynthesis(MIPPO)in the treatment of anterior pelvic ring injuries.Methods Between December 2017 and July 2023,we treated 15 patients with anterior pelvic ring injuries by using 3D printing combined with MIPPO.Before surgery,a 3D model was printed to establish the actual structure,and a steel plate was pre-bent outside the body to simulate the surgery,as well as the design of number,direction,and length of screws.Then,the pre-bent internal fixation device was inserted through the bone membrane tunnel in the anterior pelvic ring,and the screws were fixed through both sides of the incision according to the preoperative planning.Results All the 15 patients underwent surgery successfully without complications such as nerve and vascular injury.The operation time was(162.9±63.1)min,the total length of incision was(13.5±2.2)cm,the number of fluoroscopic examinations during the surgery was(5.5±1.0)times,and the amount of blood loss was(456.7±272.5)ml.The fracture healing time was(17.5±1.8)weeks.The 15 patients were followed up for(15.2±3.9)months.The Majeed functional score increased significantly from(18.3±2.3)points before surgery to(86.3±6.0)points at 6 months postoperatively(t=-47.313,P=0.000).The fracture reduction according to the Matta standards showed excellent in 14 patients,fair in one patient,the excellent and good rate being 93.3％(14/15).Conclusion Combination of 3D printing and MIPPO treatment for anterior pelvic ring injuries can achieve good clinical effects.

Objective To investigate preliminary clinical effects of three-dimensional(3D)printing combined with minimally invasive percutaneous plate osteosynthesis(MIPPO)in the treatment of anterior pelvic ring injuries.Methods Between December 2017 and July 2023,we treated 15 patients with anterior pelvic ring injuries by using 3D printing combined with MIPPO.Before surgery,a 3D model was printed to establish the actual structure,and a steel plate was pre-bent outside the body to simulate the surgery,as well as the design of number,direction,and length of screws.Then,the pre-bent internal fixation device was inserted through the bone membrane tunnel in the anterior pelvic ring,and the screws were fixed through both sides of the incision according to the preoperative planning.Results All the 15 patients underwent surgery successfully without complications such as nerve and vascular injury.The operation time was(162.9±63.1)min,the total length of incision was(13.5±2.2)cm,the number of fluoroscopic examinations during the surgery was(5.5±1.0)times,and the amount of blood loss was(456.7±272.5)ml.The fracture healing time was(17.5±1.8)weeks.The 15 patients were followed up for(15.2±3.9)months.The Majeed functional score increased significantly from(18.3±2.3)points before surgery to(86.3±6.0)points at 6 months postoperatively(t=-47.313,P=0.000).The fracture reduction according to the Matta standards showed excellent in 14 patients,fair in one patient,the excellent and good rate being 93.3％(14/15).Conclusion Combination of 3D printing and MIPPO treatment for anterior pelvic ring injuries can achieve good clinical effects.