As one of the key enzymes in cell metabolism, the activity of citrate synthase 3 (CS3) regulates the substance and energy metabolism of organisms. The protein members of CS3 family were identified from the whole genome of apple, and bioinformatics analysis was performed and expression patterns were analyzed to provide a theoretical basis for studying the potential function of CS3 gene in apple. BLASTp was used to identify members of the apple CS3 family based on the GDR database, and the basic information of CS3 protein sequence, subcellular localization, domain composition, phylogenetic relationship and chromosome localization were analyzed by Pfam, SMART, MEGA5.0, clustalx.exe, ExPASy Proteomics Server, MEGAX, SOPMA, MEME, WoLF PSORT and other software. The tissue expression and inducible expression characteristics of 6 CS3 genes in apple were determined by acid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Apple CS3 gene family contains 6 members, and these CS3 proteins contain 473-608 amino acid residues, with isoelectric point distribution between 7.21 and 8.82. Subcellular localization results showed that CS3 protein was located in mitochondria and chloroplasts, respectively. Phylogenetic analysis divided them into 3 categories, and the number of genes in each subfamily was 2. Chromosome localization analysis showed that CS3 gene was distributed on different chromosomes of apple. The secondary structure of protein is mainly α-helix, followed by random curling, and the proportion of β-angle is the smallest. The 6 members were all expressed in different apple tissues. The overall expression trend from high to low was the highest relative expression content of MdCS3.4, followed by MdCS3.6, and the relative expression level of other members was in the order of MdCS3.3 > MdCS3.2 > MdCS3.1 > MdCS3.5. qRT-PCR results showed that MdCS3.1 and MdCS3.3 genes had the highest relative expression in the pulp of 'Chengji No. 1' with low acid content, and MdCS3.2 and MdCS3.3 genes in the pulp of 'Asda' with higher acid content had the highest relative expression. Therefore, in this study, the relative expression of CS3 gene in apple cultivars with different acid content in different apple varieties was detected, and its role in apple fruit acid synthesis was analyzed. The experimental results showed that the relative expression of CS3 gene in different apple varieties was different, which provided a reference for the subsequent study of the quality formation mechanism of apple.
Citric Acid ; Malus/genetics* ; Citrate (si)-Synthase ; Phylogeny ; Citrates

Citric Acid Cycle ; Citric Acid ; Malates/metabolism* ; Citrates/metabolism* ; Aspartic Acid/metabolism*

BACKGROUND: The Coapresta 2000 (Sekisui Medical Co., Japan) is a newly developed, fully automated coagulation analyzer that can perform clotting time assays using the synthetic substrate method and the latex turbidimetric method. In this study, we evaluated the analytical performance of the Coapresta 2000 for measuring prothrombin time (PT) and activated partial thromboplastin time (aPTT), and compared the results to those of the CA-7000 (Sysmex Co., Japan) and ACL-9000 (Instrumentation Laboratory, USA) analyzers. METHODS: The Coapresta 2000 was evaluated for its precision at measuring PT and aPTT in fresh normal plasma and fresh abnormal plasma. Three hundred venous blood specimens were collected in 3.2% sodium citrate tubes, and PT and aPTT results were compared among the Coapresta 2000, ACL-9000, and CA-7000 analyzers. RESULTS: The coefficients of variation of both intra- and inter-assays for the Coapresta 2000 were <5% for PT and aPTT in the normal and pathological ranges. The results obtained using the Coapresta 2000 analyzer correlated well with those obtained using the ACL-9000 analyzer (r in the range of 0.9799-0.9886) except for aPTT (r=0.7626) and with those obtained using the CA-7000 analyzer (r in the range of 0.8258 - 0.9735). CONCLUSIONS: The Coapresta 2000 provided satisfactory precision, and the results obtained correlated well with those obtained using the existing CA-7000 and ACL-9000 coagulation analyzers. We conclude that the Coapresta 2000 would be a useful analyzer for routine coagulation tests.
Citrates ; Citric Acid ; Latex ; Partial Thromboplastin Time ; Plasma ; Prothrombin Time ; Sodium

OBJECTIVE: To evaluate the relationship between 24 h urinary ion content and kidney stones, and to explore the diagnostic values of kidney stone in primary gout patients. METHODS: Patients diagnosed with primary gout had ultrasound scanning of both feet and kidneys in Peking University First Hospital from Jan. 2020 to May 2021. Their clinical characteristics were compared between the positive and negative kidney stone groups, and the relationship between kidney stone and urinary ion composition were analyzed. Risk factors of kidney stone were analyzed. The explored diagnostic values were evaluated for urinary oxalate and citrate according with uric acid kidney stones by dual-energy computed tomography (DECT). RESULTS: Among the 100 gouty patients, 80 patients had uric acid crystal deposition in lower joints of extremity by ultrasonography, 61 patients had kidney stone, and 34 had kidney uric acid stones by DECT. All the multiple kidney stones were proved as uric acid kidney stones by DECT. Compared with patients without kidney stone group proved by ultrasonography, patients with kidney stone had longer gouty duration [(48.7±26.6) months vs. (84.0±30.6) months, P=0.01], higher 24 h urinary oxalate [(20.1±9.6) mg vs. (28.6±20.7) mg, P=0.001] and lower 24 h urinary citrate [(506.3±315.4) mg vs. (355.7±219.6) mg, P=0.001]. Compared with the patients without kidney stone by DECT, the patients with uric acid kidney stone also had longer disease duration [(49.1±28.4) months vs. (108.3±72.2) months, P=0.001], higher 24 h urinary oxalate [(23.6±16.9) mg vs. (28.5±18.8) mg, P < 0.05], lower 24 h urinary citrate [(556.0±316.3) mg vs. (391.7±261.2) mg, P < 0.05], higher serum uric acid [(466.2±134.5) μmol/L vs. (517.2±18.1) μmol/L, P < 0.05] and higher 24 h urinary uric acid [(1 518.1±893.4) mg vs. (1 684.2±812.1) mg, P < 0.05]. Logistic regression analysis showed long gout disease duration (OR=1.229, 95%CI: 1.062-1.522, P < 0.05), high serum uric acid level (OR=1.137, 95%CI: 1.001-1.213, P=0.01), low 24 h urinary citrate (OR=0.821, 95%CI: 0.659-0.952, P=0.01) were all risk factors of kidney stones by ultrasonography. Also, long gout disease duration (OR=1.201, 95%CI: 1.101-1.437, P=0.005), high serum creatine uric level (OR=1.145, 95%CI: 1.001-1.182, P=0.04), low 24 h urinary citrate (OR=0.837, 95%CI: 0.739-0.931, P=0.02) were all risk factors of kidney uric acid stones by DECT. CONCLUSION Long disease duration and low 24 h urinary citrate were risk factors for kidney stones.
Humans ; Urinary Calculi ; Uric Acid/analysis* ; Citric Acid ; Kidney Calculi/diagnostic imaging* ; Gout/diagnostic imaging* ; Citrates ; Oxalates

We report a patient with systemic lupus erythematosus (SLE), who had developed metabolic alkalosis during plasmapheresis. The metabolic alkalosis could be promptly corrected by reducing the amount of citrate load. The development of metabolic alkalosis can be explained by the citrate load during plasmapheresis. Careful monitoring of acid base status is mandatory in patients with limited renal function and the reduction of citrate load may be advisable in plasmapheresis.
Adolescent ; Alkalosis/*etiology ; Citrates ; Citric Acid ; Female ; Humans ; Lupus Erythematosus, Systemic/*metabolism/therapy ; Plasmapheresis/*adverse effects/methods

Ga-67 scintigraphy demonstrated increased uptake in the lungs and stomach in a 26-year-old man with hypercalcemia. A primitive neuroectodermal tumor was confirmed by bone marrow examination. Tc-99m MDP uptake in the same locations as Ga-67 revealed by bone scintigraphy was consistent with metastatic calcification. Although the mechanism of Ga-67 uptake in metastatic calcification is not understood, the presence of an inflammatory process is suggested.
Adult ; Bone Marrow Examination ; Citrates ; Citric Acid ; Gallium ; Humans ; Hypercalcemia ; Lung ; Neuroectodermal Tumors, Primitive ; Stomach ; Technetium Tc 99m Medronate

OBJECTIVES: Safe and effective anticoagulation is essential for hemodialysis patients who are at high risk of bleeding. The purpose of this trial is to evaluate the effectiveness and safety of two-stage regional citrate anticoagulation (RCA) combined with sequential anticoagulation and standard calcium-containing dialysate in intermittent hemodialysis (IHD) treatment. METHODS: Patients at high risk of bleeding who underwent IHD from September 2019 to May 2021 were prospectively enrolled in 13 blood purification centers of nephrology departments, and were randomly divided into RCA group and saline flushing group. In the RCA group, 0.04 g/mL sodium citrate was infused from the start of the dialysis line during blood draining and at the venous expansion chamber. The sodium citrate was stopped after 3 h of dialysis, which was changed to sequential dialysis without anticoagulant. The hazard ratios for coagulation were according to baseline. RESULTS: A total of 159 patients and 208 sessions were enrolled, including RCA group (80 patients, 110 sessions) and saline flushing group (79 patients, 98 sessions). The incidence of severe coagulation events of extracorporeal circulation in the RCA group was significantly lower than that in the saline flushing group (3.64% vs. 20.41%, P<0.001). The survival time of the filter pipeline in the RCA group was significantly longer than that in the saline flushing group ((238.34±9.33) min vs. (221.73±34.10) min, P<0.001). The urea clearance index (Kt/V) in the RCA group was similar to that in the saline flushing group with no statistically significant difference (1.12±0.34 vs. 1.08±0.34, P=0.41). CONCLUSIONS Compared with saline flushing, the two-stage RCA combined with a sequential anticoagulation strategy significantly reduced extracorporeal circulation clotting events and prolonged the dialysis time without serious adverse events.
Humans ; Citric Acid/adverse effects* ; Prospective Studies ; Sodium Citrate ; Hemorrhage/chemically induced* ; Citrates/adverse effects* ; Anticoagulants/adverse effects* ; Renal Dialysis/adverse effects*

Acute Kidney Injury/therapy* ; Anticoagulants/therapeutic use* ; Blood Coagulation ; Child ; Citrates ; Citric Acid/therapeutic use* ; Continuous Renal Replacement Therapy ; Humans ; Renal Replacement Therapy

Potassium citrate therapy caused a sustained increase in urinary pH and potassium, and restored urinary citrate to normal levels. No significant changes occurred in urinary uric acid, oxalate, sodium or phosphorus levels. Owing to these physiological changes, uric acid solubility increased, urinary saturation of calcium oxalate decreased and the propensity for spontaneous nucleation of calcium oxalate was reduced to normal. Therefore, the Physicochemical environment of urine following treatment become less conductive to the crystallization of calcium oxalate or uric acid. Twenty six patients with uric acid nephrolithiasis with or without calcium nephrolithiasis underwent treatment and long-term preventive management (mean of 20.8 months) with potassium citrate. Urinary pH increased from acid (5.0-5.5) to normal (6.5-7.0) during treatment. During the period of preventive management, stones were not developed.
Calcium ; Calcium Oxalate ; Citric Acid ; Crystallization ; Humans ; Hydrogen-Ion Concentration ; Nephrolithiasis ; Oxalic Acid ; Phosphorus ; Potassium Citrate* ; Potassium* ; Solubility ; Uric Acid ; Urinary Calculi*

PURPOSE: Citrate is a well recognized inhibitor of the formation of calcium oxalate and calcium phosphate stones. Hypocitraturia is a common etiology of recurrent calcium nephrolithiasis, with an incidence of 19 to 63%. Potassium citrate therapy can be a useful therapeutic approach for the management of calcium nephrolithiasis. But pharmacological treatment of hypocitraturic calcium nephrolithiasis requires taking too many tablets, or numerous crystal package or liquid supplements throughout the day. This cumbersome regimen often decreases patient compliance. We administered dietary citrate via lemon juice to stone former and evaluated the change of citrate levels. MATERIALS AND METHODS: The prospective study included 7 women and 8 men with documented recurrent or multiple urinary stone disease. None of the subjects suffered from renal impairment, urinary tract infection and other metabolic disorder. Controls comprised 6 voluntary men. They had no previous stone history and no evidence of stone. Patients ingested total 1 liter of lemon juice(containing 4.0gm/L.citrate) divided at 6 hours interval without strict diet restriction. Urine specimens were obtained for urinary citrate levels after 2-3days of lemon juice therapy and compared to pre-lemon juice baseline values. RESULTS: All 15 patient showed increased urinary citrate levels during lemon juice therapy. Average urinary citrate levels increased from 146+/-109mg/day at baseline to 453+/-226mg/day during treatment(p<0.05). Urinary citrate levels during treatment increased up to those of control group(351+/-265mg/day) and did not show significant difference (p>0.05). Urinary pH increased from 5.9+/-0.4 at baseline to 6.8+/-0.6 during treatment(p<0.05). No patient complained of gastrointestinal discomforts. CONCLUSIONS: Citrate supplementation with lemon juice increased urinary citrate levels and urinary pH. Lemon juice is well tolerated dietary source of citrate and would be beneficial in the control of calcium urolithiasis.
Calcium ; Calcium Oxalate ; Citric Acid* ; Diet ; Female ; Humans ; Hydrogen-Ion Concentration ; Incidence ; Male ; Nephrolithiasis ; Patient Compliance ; Potassium Citrate ; Prospective Studies ; Tablets ; Urinary Calculi ; Urinary Tract Infections ; Urolithiasis*