OBJECTIVES: To analyze the differentially expressed exosomal miRNAs in patients with chronic heart failure (CHF) complicated by hyperuricemia (HUA) and explore their potential as novel diagnostic molecular markers and their target genes. METHODS: This study was conducted among 30 CHF patients with HUA (observation group) and 30 healthy volunteers (control group) enrolled between September, 2020 and September, 2023. Peripheral blood samples were collected from 6 CHF patients with HUA for analyzing exosomal miRNAs by high-throughput sequencing, and the results were validated in the remaining 24 patients using qRT-PCR. GO and KEGG enrichment analyses were performed to predict the the target genes of the identified differential miRNAs. We also validated the differentially expressed miRNAs by animal experiment. RESULTS: A total of 42 differentially expressed exosomal miRNAs were detected in observation group by high-throughput sequencing; among them, miR-27a-5p was significantly upregulated (P=0.000179), and miR-139-3p was significantly downregulated (P=0.000058). In the 24 patients with both CHF and PUA, qRT-PCR validated significant upregulation of miR-27a-5p (P=0.004) and downregulation of miR-139-3p (P=0.005) in serum exosomes. When combined, miR-27a-5p and miR-139-3p had a maximum area under the curve (AUC) of 0.899 (95% CI: 0812-0.987) for predicting CHF complicated by HUA. GO and KEGG enrichment analyses suggested that the differential expressions of miR-27a-5p and miR-139-3p was associated with the activation of the AMPK-mTOR signaling pathway to activate the autophagic response. We obtained the same conclusion from animal experiment. CONCLUSIONS Upregulated exosomal miR-27a-5p combined with downregulated exosomal miR-139-3p expression can serve as a novel molecular marker for diagnosis of CHF complicated by HUA, and their differential expression may promote autophagy in cardiomyocytes by activating the AMPK-mTOR signaling pathway.
Humans ; Hyperuricemia/diagnosis* ; Heart Failure/genetics* ; MicroRNAs/metabolism* ; Exosomes/metabolism* ; Chronic Disease ; Male ; Female ; Middle Aged ; Animals

This study aims to investigate the biological function of the Clumping factor B(clfB)gene in Staphylococcus aureus.The recombinant plasmid pBT2-△clfB was constructed and elec-troporated into Staphylococcus aureus J57 to delete clfB by homologous recombination.The ex-pression plasmid pLI50-clfB was constructed,modified,and electroporated into clfB gene deletion strain △clfB and constructed a complementation strain(C△clfB).J57,△clfB,and C△clfB were cultured at 37℃ for 12 h,and the growth curves of each strain were plotted.The hemolytic properties of each strain were analyzed by contact method,the motility of each strain on TSA plates was determined,and the autolysis rate of each strain under the action of TritonX-100 was determined.Crystal violet staining was used to detect each strain's biofilm formation ability,and biofilm components formed by each strain were quantitatively analyzed.The K-B method was used to determine the sensitivity of each strain to commonly used antibiotics.ClfB gene deletion strain△clfB and the complemented strain C△clfB were successfully constructed.The growth curves of the deletion strain were almost consistent with those of the wild and complemented strains,and there was no significant difference.Compared with J57 and C△clfB,the hemoly ability and the athletic of △clfB decreased.In the condition of TritonX-100,the autolysis rate of △clfB was significantly lower than that of J57 and C△clfB(P＜0.01).Compared with J57 and C△clfB,the ability of △clfB to form biofilm was significantly lower than that of J57 and C△clfB(P＜0.05),and the content of extracellular DNA and protein in the biofilm was significantly decreased.In con-trast,the content of soluble polysaccharides was significantly increased(P＜0.05).Compared with J57 and C△clfB,△clfB was more sensitive to chloramphenicol,gentamicin,and kanamycin while more resistant to linezolid.ClfB gene is closely related to the autolysis,hemolytic activity,athlet-ic,and biofilm formation ability of Staphylococcus aureus,affecting the sensitivity of bacteria to certain antibiotics.

This study aims to investigate the biological function of the Clumping factor B(clfB)gene in Staphylococcus aureus.The recombinant plasmid pBT2-△clfB was constructed and elec-troporated into Staphylococcus aureus J57 to delete clfB by homologous recombination.The ex-pression plasmid pLI50-clfB was constructed,modified,and electroporated into clfB gene deletion strain △clfB and constructed a complementation strain(C△clfB).J57,△clfB,and C△clfB were cultured at 37℃ for 12 h,and the growth curves of each strain were plotted.The hemolytic properties of each strain were analyzed by contact method,the motility of each strain on TSA plates was determined,and the autolysis rate of each strain under the action of TritonX-100 was determined.Crystal violet staining was used to detect each strain's biofilm formation ability,and biofilm components formed by each strain were quantitatively analyzed.The K-B method was used to determine the sensitivity of each strain to commonly used antibiotics.ClfB gene deletion strain△clfB and the complemented strain C△clfB were successfully constructed.The growth curves of the deletion strain were almost consistent with those of the wild and complemented strains,and there was no significant difference.Compared with J57 and C△clfB,the hemoly ability and the athletic of △clfB decreased.In the condition of TritonX-100,the autolysis rate of △clfB was significantly lower than that of J57 and C△clfB(P＜0.01).Compared with J57 and C△clfB,the ability of △clfB to form biofilm was significantly lower than that of J57 and C△clfB(P＜0.05),and the content of extracellular DNA and protein in the biofilm was significantly decreased.In con-trast,the content of soluble polysaccharides was significantly increased(P＜0.05).Compared with J57 and C△clfB,△clfB was more sensitive to chloramphenicol,gentamicin,and kanamycin while more resistant to linezolid.ClfB gene is closely related to the autolysis,hemolytic activity,athlet-ic,and biofilm formation ability of Staphylococcus aureus,affecting the sensitivity of bacteria to certain antibiotics.

Animals ; Antigens, Surface/genetics* ; Cell Communication/genetics* ; Copulation/physiology* ; Fallopian Tubes/metabolism* ; Female ; Fertilization/genetics* ; GPI-Linked Proteins/genetics* ; Gene Expression Regulation ; Genes, Reporter ; Green Fluorescent Proteins/metabolism* ; Litter Size ; Luminescent Proteins/metabolism* ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mitochondria/metabolism* ; Reproduction/genetics* ; Signal Transduction ; Sperm Count ; Sperm Motility/genetics* ; Spermatozoa/metabolism* ; Uterus/metabolism*

Objective: To investigate the urine methyl tert-butyl ether (MTBE) level of a primary school students in Changping District, Beijing, and analysis the effect factors, give the advice if the MTBE is eligibility as an exposure biomarker of automobile exhaust. Methods: Using the panel study, randomly selected 20 sixthgrade students (10 male and 10 female) from Zhixing School in Changping district, Beijing, from June 7th to 13th, 2018. All participants completed a questionnaire for collecting demographic and geographic information and daily activities. All participants were given guidance to repeatedly collected urine samples 3 times (8:00, 11:00 and 16:00 instant urien) a day for 7 days, totally 420 urine samples were collected. Urine MTBE were measured and the statistic significant of the Urine MTBE at different groups were evaluated. Results: MTBE were detected in all 420 urine samples of primary school students, the geometric mean of creatinine-corrected MTBE exposure level in 8:00, 11:00 and 16:00 instant urine were 0.12, 0.12 and 0.12 mg/g creatinine, respectively. Urine MTBE concentration was statistical significant in different daily outdoor activity time, the distance of home address to the main road, and daily use time of air purification equipment in class(P=0.00, 0.04, 0.00). Conclusion There was a universal MTBE exposure among students in a primary school in Beijing, and the impact factors to exposure level were daily outdoor activity time, distance from home address to the closest main road, and daily use time of air purification equipment in class. This result indicate MTBE may be a valuable candidate for exposure biomarker of automobile exhaust, and more research should be done.

Objective To explore the effect of airway humidification on lung injury as a result of mechanical ventilation with different tidal volume(VT). Methods Twenty-four male Japanese white rabbits were randomly divided into four groups:low VT with airway humidification group,high VT with airway humidification group,low VT and high VT group without humidification,with 6 rabbits in each group. Mechanical ventilation was started after intubation and lasted for 6 hours. Low VT denoted 8 mL/kg,while high VT was 16 mL/kg,fraction of inspired oxygen (FiO2)denoted 0.40,positive end-expiratory pressure(PEEP)was 0. Temperature at Y piece of circuit in airway humidification groups was monitored and controlled at 40℃. Arterial blood gas analysis,including pH value,arterial partial pressure of oxygen(PaO2),arterial partial pressure of carbon dioxide(PaCO2),lung mechanics indexes, including peak airway pressure(Ppeak)and airway resistance(Raw),and lung compliance was measured at 0,2,4, 6 hours of mechanical ventilation. The levels of tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)in plasma and bronchoalveolar lavage fluid(BALF)were determined by enzyme linked immunosorbent assay(ELISA). The animals were sacrificed at the end of mechanical ventilation. The wet to dry(W/D)ratio of lung tissues was calculated. Histopathologic changes in the lung tissueies were observed with microscope,and lung injury score was calculated. Scanning and transmission electron microscopies were used to examine the integrity of the airway cilia and the tracheal epithelium. Results Compared with low VT group,pH value in high VT group was significantly increased,PaCO2 was significantly lowered,and no difference in PaO2 was found. Ppeak,Raw,and lung compliance were significantly increased during mechanical ventilation. There were no significant differences in blood gas analysis and lung mechanics indexes between low VT with airway humidification group and low VT group. Compared with high VT group,PaCO2 in high VT with airway humidification group was significantly decreased,Ppeak raised obviously,and no difference in pH value,PaO2,Raw and pulmonary compliance was found. Compared with low VT with airway humidification group,no difference in blood gas analysis(PaCO2,mmHg,1 mmHg=0.133 kPa)was found,but Ppeak(cmH2O,1 cmH2O=0.098 kPa),Raw(cmH2O),and lung compliance(mL/cmH2O)were increased significantly in high VT with airway humidification group(PaCO2 at 2 hours:27.96±4.64 vs. 36.08±2.11,4 hours:28.62±2.93 vs. 34.55±5.50, 6 hours:29.33±2.14 vs. 35.01±5.53;Ppeak at 0 hour:14.34±1.97 vs. 8.84±1.32,2 hours:17.33±0.52 vs. 11.17±2.14,4 hours:17.83±0.98 vs. 12.67±2.06,6 hours:18.67±1.22 vs. 13.50±2.16;Raw at 0 hour:37.36±5.14 vs. 27.05±2.93,2 hours:43.94±6.58 vs. 31.95±3.56,4 hours:48.04±6.07 vs. 35.24±3.50, 6 hours:50.33±6.34 vs. 36.66±3.64;pulmonary compliance at 6 hours:2.28±0.18 vs. 1.86±0.37,all P<0.05). The lung W/D ratio in high VT group was significantly higher than that of the low VT group(6.17±2.14 vs. 3.50±1.52, P<0.05). W/D in high VT with airway humidification group was higher than that of low VT with airway humidification group but without statistically significant difference(5.17±2.14 vs. 3.00±1.10,P>0.05). Microscopic observation showed that cilia were partially detached,adhered and sparse in low VT group,while cilia in high VT group showed serious detachment and lodging. Remaining cilia were sparse,with lodging,and cellular structure was damaged. Lung tissue pathological injury score in the high VT group was significantly higher than that of low VT group(6.17±2.14 vs. 3.50±1.52,P<0.05). Cilia density and cellularity were normal in low VT with airway humidification group,and no difference in lung tissue pathological injury score was found compared with low VT group(3.00±1.10 vs. 3.50±1.52, P>0.05). Cilia were severely detached,adhered and lodging,and cellularity were not obvious in high VT with airway humidification group,and lung tissue pathological injury score was elevated significantly than that of the low VT with airway humidification group but without statistically significant difference(5.17±2.14 vs. 3.00±1.10,P>0.05). TNF-α and IL-8 concentrations showed no change in plasma and BALF in all groups during ventilation,and no significant difference was found among the groups. Conclusions Airway humidification can alleviate pathological lung injury,damage of cilia and cellular structure in trachea caused by mechanical ventilation with low and high VT. High VT with humidification can result in serious pulmonary edema.

OBJECTIVE:To analysis outcomes of patients with knee osteoarthritis treated by mini-incision knee replacement accompanied by early rehabilitation. METHODS:Outcomes of 28 knee osteoarthritis patients (46 knees) who were treated by joint replacement from 2005 June to August were retrospectively analyzed. Sixteen patients were served as experimental group,including 4 males and 12 females,aged 59-77 (65.3?5.3) years,and the course of disease was 10-15 (13.4?4.3) years. These patients were received mini-incision knee replacement,accompanied by specified early rehabilitation procedure. Early rehabilitation procedure comprised psychological treatment,training before knee replacement,muscle strength training after knee replacement,and correction of poor gait,joint position sense training and exercise for coordination after knee replacement,early quadriceps femoral muscle active extension knee exercises especially emphasized. Another 12 patients were served as control group,including 5 males and 7 females,aged 60-75 (67.5?4.7) years,with course of disease 8-18 (12.8?4.9) years,they were received normal knee replacement. Pain severity,joint motion ranges and functional situations were evaluated. RESULTS:Twenty-eight patients stayed in hospital for (13.2?2.7) days (ranging from 10 days to 16 days). All the patients were followed up 9 to 12 months. There were no significantly differences in pain severity,joint motion ranges and functional scores on admission between two groups. Pain,joint motion and joint functions were improved greatly after knee replacement. The joint motion ranges and functional scores were higher on discharge and the last follow-up in the experimental group than that of the control group. CONCLUSION:The program that consists of Mini-incision knee replacement,which protected quadricep muscle intactness and diminished surgical injury,accompanied by psychological treatment,preoperational preparation,postoperational training,correction of gait,training in articular position sensation and all the body coordination,can improve the outcomes of the knee osteoarthritis patients with knee replacement.