Objective To investigate the effect and mechanism of Efferocytosis Relatived LncRNA (EFRL) on homocysteine-induced atherosclerosis in macrophage efferocytosis. Methods RAW264.7 cells were cultured in vitro, and the Control group (0 μmol/L Hcy) and Hcy intervention group (100 μmol/L Hcy) were set up. After GapmeR transfection of macrophages with Hcy intervention, EFRL knockdown negative control group (Hcy combined with LNA-NC) and EFRL knockdown group (Hcy combined with LNA-EFRL) were set up. High-throughput sequencing was applied for different expression of LncRNA MSTRG. 88917.16 (EFRL), UCSC was used to analyze its conservation, CPC and CPAT were used to analyze its ability to encode proteins, and GO and KEGG were used to analyze related biological functions. The localization of LncRNA EFRL in macrophages was analyzed by nucleoplasmic separation and RNA-FISH. Quantitative real-time PCR was used to detect the expression levels of LncRNA EFRL and its target gene SPAST in Hcy-treated macrophages. The apoptosis rate of Jurkat cells induced by UV was detected by flow cytometry. In vitro efferocytosis assay combined with immunofluorescence technique was used to analyze macrophage efferocytosis. ELISA was used to detect the levels of interleukin 1β(IL-1β) and IL-18. Results The new LncRNA MSTRG.88917.16 was identified and named EFRL(Efferocytosis Relatived LncRNA). UCSC, CPC and CPAT analyses showed that LncEFRL is highly conserved and does not have the ability to encode proteins. GO and KEGG analyses suggested that LncEFRL may be involved in macrophage efferocytosis. LncRNA EFRL was localized in the nucleus of macrophages as determined by nucleoplasmic separation and RNA-FISH. In comparison to the Control group, the expression levels of LncRNA EFRL and its target gene SPAST in the Hcy group were increased. In comparison to the Control group (0 min), the apoptosis rate of the experimental group (15, 30 min) Annexin V is more than 85%. Compared with Hcy combined with LNA-NC group, Hcy combined with LNA-EFRL group had enhanced macrophage efferocytosis and reduced levels of inflammatory factors. Compared with Hcy combined with LNA-NC group, the expression level of SPAST in Hcy combined with LNA-EFRL group was decreased. Conclusion Inhibition of EFRL expression can alleviate the process of Hcy inhibiting macrophage efferocytosis, and the mechanism is related to the regulation of the downstream target gene SPAST by EFRL.
RNA, Long Noncoding/physiology* ; Animals ; Homocysteine ; Mice ; Macrophages/drug effects* ; Humans ; RAW 264.7 Cells ; Atherosclerosis/chemically induced* ; Apoptosis/genetics* ; Phagocytosis/genetics* ; Jurkat Cells ; Interleukin-1beta/genetics* ; Efferocytosis

This study aims to explore the correlation between plasma homocysteine (Hcy) levels and the clinical grading of varicocele (VC) when analyzing the potential pathogenesis of endothelial cells injury by Hcy. A total of 184 VC patients, aged 18-46 years, were included in this study. These patients visited The Second Hospital of Dalian Medical University (Dalian, China), between January 2022 and September 2024. Patients were divided into three groups based on clinical grading: Group A (59 cases, Grade I), Group B (28 cases, Grade II), and Group C (97 cases, Grade III). Additionally, 120 individuals with normal fertility test results during the same period were selected as the control group. Routine blood and biochemical indices were collected from the patients. Differences in clinical indices between groups were compared, and univariate and multivariate linear regression analyses were performed to identify factors associated with clinical grading. The results showed that the median Hcy levels in the control group and in patients with Grade I, II, and III VC were 9.56 (interquartile range [IQR]: 8.66, 14.02) µmol l -1 , 11.28 (IQR: 9.71, 14.55) µmol l -1 , 11.84 (IQR: 10.14, 15.60) µmol l -1 , and 12.27 (IQR: 9.52, 15.40) µmol l -1 , respectively. The differences between the four groups were statistically significant ( χ2 = 12.41, P = 0.006). Multivariate regression analysis indicated that Hcy is a factor associated with the clinical grading of VC ( t = 2.53, P = 0.013). Hcy is associated with the clinical grading and may have clinical value in assessing severity of VC.
Humans ; Varicocele/pathology* ; Male ; Homocysteine/blood* ; Adult ; Middle Aged ; Adolescent ; Young Adult ; Severity of Illness Index ; Case-Control Studies

OBJECTIVE: Data on homocysteine (Hcy) status and its determinants are limited among women during pregnancy and postpartum. This cross-sectional study aimed to investigate Hcy levels during pregnancy and postpartum, and to explore the determinants like geographic factor. METHODS: This study was conducted in women at mid-pregnancy, late-pregnancy and postpartum from southern, central and northern China. Approximately 132 women were included in each stratum by the three phases and regions. Plasma Hcy concentrations were assessed using high-performance liquid chromatography (HPLC), with hyperhomocysteinemia defined as > 10.0 µmol/L. Quantile regression was to estimate medians and interquartile ranges ( IQRs), and logistic regression to examine the determinants of hyperhomocysteinemia. RESULTS: For 1,190 women included, the median (IQR) Hcy concentration was 5.66 (4.62, 7.37) μmol/L. The adjusted median in mid-pregnancy, late-pregnancy and postpartum women was 4.75 (4.13, 5.54), 5.72 (4.81, 6.85) and 7.09 (5.65, 8.75) μmol/L, respectively, showing an increasing trend ( P < 0.001). This increasing trend persisted across the three regions. Higher Hcy concentrations were observed in women residing in northern region and those with younger age or lower economic status. A total of 106 (8.9%) women had hyperhomocysteinemia, with a higher prevalence in those residing in northern region (16.0%), or in postpartum women (16.5%). CONCLUSION Hcy levels, varying with geographic region, maternal age and economic status, are increased from mid-pregnancy to late-pregnancy and postpartum, indicating a need to monitor Hcy levels in pregnant and postpartum women to control potential risks related to elevated Hcy levels.
Humans ; Female ; Pregnancy ; Homocysteine/blood* ; China/epidemiology* ; Adult ; Postpartum Period/blood* ; Cross-Sectional Studies ; Hyperhomocysteinemia/blood* ; Young Adult ; Pregnancy Trimester, Third/blood* ; Pregnancy Trimester, Second ; East Asian People

Objective To investigate the association of the serum levels of ischemia-modified albumin（IMA）and homocysteine（Hcy）with the onset and severity of white matter ischemic lesions（WMIL）. Methods A total of 89 patients who were diagnosed with WMIL by cranial magnetic resonance imaging（MRI）in our hospital from March 2020 to June 2022 were enrolled as WMIL group，and the volunteers with normal cranial MRI results，matched for sex and age，during the same period of time were enrolled as control group. The two groups were compared in terms of general information and laboratory markers，and the conditions of WMIL were evaluated and classified into mild and moderate-to-severe cases. A logistic regression analysis was used to investigate the influencing factors for the onset and severity of WMIL，and the receiver operating characteristic（ROC）curve was used to analyze the value of the serum levels of IMA and Hcy in the diagnosis of WMIL. Results Compared with the control group，the WMIL group had a significantly higher proportion of patients with hypertension or type 2 diabetes and significantly higher levels of uric acid（UA），IMA，and Hcy（P<0.05）. Compared with the control group，the WMIL group had a significantly higher proportion of the patients with moderate-to-severe WMIL who had hypertension or the proportion of type 2 diabetes，significantly higher levels of IMA and Hcy，and a significantly lower level of high-density lipoprotein cholesterol（P<0.05）. The logistic regression analysis showed that hypertension，elevated IMA level，and elevated Hcy level were influencing factors for the onset of WMIL（P<0.05），and hypertension，elevated IMA level，and elevated Hcy level were influencing factors for the increase in WMIL severity（P<0.05）. The ROC curve analysis showed that serum IMA and Hcy had a certain value in the diagnosis of WMIL and moderate-to-severe WMIL. Conclusion Increases in the serum levels of IMA and Hcy are associated with the onset and aggravation of WMIL，and measurement of the serum levels of IMA and Hcy has a certain value in the diagnosis of WMIL and moderate-to-severe WMIL.
Homocysteine

Objective To explore the value of serum homocysteine​（HCY） and cystatin C （Cys-C） levels for evaluating the presence and severity of cognitive impairment in elderly patients with both cerebral small vessel disease （CSVD） and obstructive sleep apnea-hypopnea syndrome （OSAHS）.Methods A retrospective study was performed on 150 elderly patients with both CSVD and OSAHS who were treated in our hospital from January to December 2020. The data about HCY and Cys-C levels were collected. According to the presence or absence of cognitive impairment， they were divided into observation group （with cognitive impairment， 41 cases） and control group （without cognitive impairment， 109 cases）. The serum HCY and Cys-C levels of the two groups were compared. The correlations between serum HCY and Cys-C levels and cognitive impairment in patients with both CSVD and OSAHS were analyzed. A binary Logistic regression analysis was performed to determine independent factors affecting cognitive impairment to establish a predictive logistic regression model. Receiver operating characteristic （ROC） curves were plotted to examine the predictive value of HCY， Cys-C， and their combination for cognitive impairment in patients with both CSVD and OSAHS. According to the severity of cognitive impairment， the patients in the observation group were further divided into mild cognitive impairment group （32 cases） and severe cognitive impairment group （9 cases） to compare HCY and Cys-C levels and analyze the value of HCY， Cys-C， and their combination in evaluating the staging of cognitive impairment by using ROC curves.Results The levels of HCY and Cys-C in the observation group were significantly higher than those in the control group （both P<0.05）. The total score of Montreal Cognitive Assessment was negatively correlated with serum HCY and Cys-C level （r=-0.756， -0.713， both P<0.001）. The logistic regression analysis showed that HCY and Cys-C levels were independent influencing factors for cognitive impairment （both P<0.05）. The area under the curve （AUC） of HCY plus Cys-C for prediction was 0.843（0.747-0.934）， which was higher than that of any single indicator （P < 0.05）， with the sensitivity and specificity being 89.32% and 79.25%， respectively. The levels of HCY and Cys-C were significantly lower in the patients with mild cognitive impairment than in those with severe cognitive impairment （both P < 0.05）. The AUC of HCY plus Cys-C in cognitive impairment staging was 0.925（0.849-0.976）， which was higher than that of any single index （P < 0.05）. The sensitivity and specificity were 95.67% and 89.67%， respectively.Conclusion Serum HCY and Cys-C levels are negatively correlated with the development and progression of cognitive impairment in elderly patients with both CSVD and OSAHS. The combined prediction with the two indicators can facilitate the evaluation of the risk and stage of cognitive impairment.
Homocysteine ; Cystatins

Objective To investigate the risk factors for the early progression of atherosclerosis，whether hyperhomocysteinemia （HHcy） is an independent risk factor for the early progression of atherosclerosis， and whether the reduction in homocysteine （Hcy） can delay the early progression of atherosclerosis. Methods The villagers，aged ≥40 years， from Liulin Town of Hanzhong City， Shaanxi Province， China were selected for structured inquiry， carotid ultrasound examination， and blood test in May 2017 and June 2020. Ultrasound examination was performed to measure carotid intima-media thickness （CIMT）， carotid plaque， and carotid stenosis rate to reflect the degree of arteriosclerosis. Results A total of 500 individuals were included in the final analysis， among whom 451 had no CIMT thickening at baseline， 429 had no carotid plaque at baseline， and 454 had no carotid stenosis at baseline. After 3 years of follow-up， among the 500 individuals， 176 （35.2%） had CIMT thickening， and the multivariate analysis showed that old age， smoking， and a high level of low-density lipoprotein cholesterol （LDL-C） were independent risk factors for CIMT thickening； 252 （50.4%） had new-onset plaques， and the multivariate analysis showed that old age， hypertension， and high LDL-C were independent risk factors for plaque formation； 231（46.2%） had new-onset carotid stenosis， and the multivariate analysis showed that old age， high LDL-C， and elevated triglyceride were independent risk factors for carotid stenosis. There were 364 patients with HHcy （≥15 μmol/L） in the initial examination， and after 3 years， 119 had a reduction in Hcy， while 245 had no reduction in Hcy. After adjustment for sex， previous stroke， smoking， baseline LDL-C， baseline Hcy， and Hcy after 3 years， the multivariate analysis showed that neither HHcy nor the reduction in Hcy was associated with carotid atherosclerosis. Conclusion Old age， high LDL-C， smoking， and hypertension are the main risk factors for atherosclerosis in villagers aged ≥40 years in rural areas of Hanzhong City in Shaanxi Province. There is no significant association between HHcy and carotid atherosclerosis， and the reduction in Hcy cannot delay or reverse the process of carotid atherosclerosis.
Atherosclerosis ; Homocysteine

Objective: By analyzing the prevalence and influencing factors of thyroid nodules (TN) among a population undergoing physical examinations in Nantong region, this study aims to provide theoretical basis for early prevention and intervention of TN. Methods: A cross-sectional study was conducted, including 6 950 participants who underwent physical examinations at the Affiliated Hospital of Nantong University from January 2017 to April 2020. All participants underwent high-resolution ultrasound examination of the thyroid, and measurements of height, body mass index (BMI), blood pressure. Fasting blood glucose (FBG), total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C), uric acid (UA), homocysteine (HCY) and other metabolic indicators were detected. Data analysis was performed using SPSS 26.0 statistical software. The numerical variables with normal distribution were expressed as mean±standard deviation (x¯±s), and the t-test was used for comparison between the two groups. Numerical variables with non-normal distribution were expressed as median (interquartile range), namely M (Q₁, Q₃). The Mann-Whitney U test was used for comparison between two groups, and the Kruskal-Wallis test was used for comparison between multiple groups. Results: The prevalence of thyroid nodules among the 6 950 participants was 53.97% (3 751/6 950), with a rate of 47.08% (2 218/4 711) in males and 68.47% (1 533/2 239) in females, which was significantly higher in females than in males (χ²=278.575, P<0.001). The prevalence of TN increased with age both overall (χ²=552.145, P<0.001), in males (χ²=304.086, P<0.001), and in females (χ²=202.178, P<0.001). The prevalence of TN was higher in females than in males across different age groups (P<0.05). In the comparison between males in the TN and non-TN groups, significant differences were found in terms of alcohol consumption history, BMI, blood pressure, HCY, and FBG (all P<0.05). In the comparison between females in the TN and non-TN groups, significant differences were found in terms of BMI, blood pressure, HCY, FBG, TC, TG, LDL-C, and UA (all P<0.05). Univariate logistic regression model showed that FBG<6.1 mmol/L (P<0.001) and TC<5.2 mmol/L (P=0.013) were protective factors for TN. Normal UA (P=0.013) was a risk factor for TN. After adjusting for gender, smoking, alcohol consumption, BMI, and blood pressure, multivariate logistic regression analysis revealed that FBG<6.1 mmol/L (OR: 0.713, 95%CI: 0.621-0.817, P<0.001) was a protective factor against TN. Conclusion: The prevalence of TN is relatively high in the Nantong region. Gender, age, blood pressure, BMI, and FBG are important influencing factors for TN. Health screening and management should be strengthened for the physical examination population with abnormal indicators.
Female ; Male ; Humans ; Thyroid Nodule/epidemiology* ; Cholesterol, LDL ; Cross-Sectional Studies ; Physical Examination ; Body Mass Index ; Homocysteine

Objective: To characterize the relationship between the levels of plasma methyl donor and related metabolites (including choline, betaine, methionine, dimethylglycine and homocysteine) and fetal growth in twin pregnancies. Methods: A hospital-based cohort study was used to collect clinical data of 92 pregnant women with twin pregnancies and their fetuses who were admitted to Peking University Third Hospital from March 2017 to January 2018. Fasting blood was collected from the pregnant women with twin pregnancies (median gestational age: 18.9 weeks). The levels of methyl donors and related metabolites in plasma were quantitatively analyzed by high-performance liquid chromatography combined with mass spectrometry. The generalized estimation equation was used to analyze the relationship between maternal plasma methyl donors and related metabolites levels and neonatal outcomes of twins, and the generalized additive mixed model was used to analyze the relationship between maternal plasma methyl donors and related metabolites levels and fetal growth ultrasound indicators. Results: (1) General clinical data: of the 92 women with twin pregnancies, 66 cases (72%) were dichorionic diamniotic (DCDA) twin pregnancies, and 26 cases (28%) were monochorionic diamniotic (MCDA) twin pregnancies. The comparison of the levels of five plasma methyl donors and related metabolites in twin pregnancies with different basic characteristics showed that the median levels of plasma choline and betaine in pregnant women ≥35 years old were higher than those in pregnant women <35 years old, and the differences were statistically significant (all P<0.05). (2) Correlation between plasma methyl donor and related metabolites levels and neonatal growth indicators: after adjusting for confounding factors, plasma homocysteine level in pregnant women with twins was significantly negatively correlated with neonatal birth weight (β=-47.9, 95%CI:-94.3- -1.6; P=0.043). Elevated methionine level was significantly associated with decreased risks of small for gestational age infants (SGA; OR=0.5, 95%CI: 0.3-0.9; P=0.021) and low birth weight infants (OR=0.6, 95%CI: 0.4-0.9; P=0.020). Increased homocysteine level was associated with increased risks of SGA (OR=1.5, 95%CI: 1.0-2.2; P=0.029) and inconsistent growth in twin fetuses (OR=1.9, 95%CI: 1.0-3.7; P=0.049). (3) Correlation between the levels of plasma methyl donors and related metabolites and intrauterine growth indicators of twins pregnancies: for every 1 standard deviation increase in plasma choline level in pregnant women with twin pregnancies, fetal head circumference, abdominal circumference, femoral length and estimated fetal weight in the second trimester increased by 1.9 mm, 2.6 mm, 0.5 mm and 20.1 g, respectively, and biparietal diameter, abdominal circumference and estimated fetal weight increased by 0.7 mm, 3.0 mm and 38.4 g in the third trimester, respectively, and the differences were statistically significant (all P<0.05). (4) Relationship between plasma methyl donor and related metabolites levels in pregnant women with different chorionicity and neonatal birth weight and length: the negative correlation between plasma homocysteine level and neonatal birth weight was mainly found in DCDA twin pregnancy (β=-65.9, 95%CI:-110.6- -21.1; P=0.004). The levels of choline, betaine and dimethylglycine in plasma of MCDA twin pregnancy were significantly correlated with the birth weight and length of newborns (all P<0.05). Conclusion: Homocysteine level is associated with low birth weight in twins, methionine is associated with decreased risk of SGA, and choline is associated with fetal growth in the second and third trimesters of pregnancy.
Adult ; Female ; Humans ; Infant, Newborn ; Pregnancy/metabolism* ; Betaine/metabolism* ; Birth Weight/physiology* ; Choline/metabolism* ; Cohort Studies ; Fetal Development/physiology* ; Fetal Weight/physiology* ; Homocysteine/metabolism* ; Methionine/metabolism* ; Pregnancy, Twin/physiology* ; Biomarkers/metabolism* ; Pregnancy Trimesters/physiology* ; Pregnancy Outcome

Objective: To investigate the relationship between subchorionic hematoma (SCH) and coagulation status, autoantibodies, and conception method. Methods: A total of 100 pregnant women diagnosed with SCH from June 2020 to December 2021 in the Third Affiliated Hospital of Zhengzhou University were included in the SCH group, while 100 healthy pregnant women during the same period were selected as the control group. The coagulation status (including platelet, prothrombin time, thrombin time, activated partial thromboplastin time, fibrinogen, antithrombin Ⅲ, fibrin degradation products, D-dimer, homocysteine, protein S activity, protein C activity), the positive rate of autoantibodies [including antiphospholipid antibodies (anticardiolipin antibody and anti-β₂ glycoprotein Ⅰ antibody), antinuclear antibody] and the mode of conception of the two groups were analyzed. Results: Compared to the control group, the SCH group had higher levels of platelet [(240±45)×10⁹/L vs (227±37)×10⁹/L], fibrinogen [(4.0±0.8) vs (3.6±0.7) g/L], D-dimer [(0.42±0.18) vs (0.31±0.15) mg/L], blood homocysteine [(8.9±4.2) vs (6.9±2.3) μmol/L], and lower level of protein S activity [(55±14)% vs (68±20)%], and there were significant differences between the two groups (all P<0.05). The SCH group had higher positive rates of autoantibodies [24.0% (24/100) vs 8.0% (8/100)], antiphospholipid antibodies [15.0% (15/100) vs 6.0% (6/100)], anti-β₂ glycoprotein Ⅰ antibody [10.0% (10/100) vs 3.0% (3/100)], antinuclear antibody [11.0% (11/100) vs 2.0% (2/100)] and assisted reproduction rate [10.0% (10/100) vs 2.0% (2/100)] than those of the control group (all P<0.05). Conclusion: The occurrence of SCH is related to blood hypercoagulability, positive autoantibodies, and assisted reproduction.
Pregnancy ; Female ; Humans ; Autoantibodies ; Antibodies, Antinuclear ; Antibodies, Antiphospholipid ; Fibrinogen ; Homocysteine ; Glycoproteins

OBJECTIVES: Hyperhomocysteinaemia (Hcy) is an independent risk factor for cardiovascular and cerebrovascular diseases. MicroRNA (miR)-18a-5p is closely related to cardiovascular diseases. This study aims to investigate the effects of miR-18a-5p on homocysteine (Hcy)-induced myocardial cells injury. METHODS: H9c2 cells were transfected with miR-18a-5p mimic/miR-18a-5p mimic negative control (NC) or combined with Hcy for intervention, and untreated cells were set as a control group. The transfection efficiency was verified by real-time RT-PCR, and cell counting kit-8 (CCK-8) assay was used to determine cell viability. Flow cytometry was used to detect apoptosis and reactive oxygen species (ROS) levels. Western blotting was performed to measure the protein levels of microtubule-associated protein 1 light chain 3 (LC3)-I, LC3-II, Beclin1, p62, Bax, Bcl-2, and Notch2. Dual luciferase reporter assay was used to detect the interaction of miR-18a-5p with Notch2. RESULTS: Compared with the control, treatment with Hcy or transfection with miR-18a-5p mimic alone, or combined treatment with Hcy and miR-18a-5p mimic/miR-18a-5p mimic NC significantly reduced the H9c2 cell viability, promoted apoptosis and ROS production, up-regulated the expressions of Bax and Beclin, down-regulated the expressions of Bcl-2, p62, and Notch2, and increased the ratio of LC3-II/LC3-I (all P<0.05). Compared with the combined intervention of miR-18a-5p mimic NC and Hcy group, the above indexes were more significantly changed in the combined intervention of miR-18a-5p mimic and Hcy group, and the difference between the 2 groups was statistically significant (all P<0.05). There is a targeted binding between Notch2 and miR-18a-5p. CONCLUSIONS MiR-18a-5p could induce autophagy and apoptosis via increasing ROS production in cardiomyocytes, and aggravate Hcy-induced myocardial injury. Notch2 is a target of miR-18a-5p.
Apoptosis/genetics* ; Autophagy/genetics* ; bcl-2-Associated X Protein ; MicroRNAs/metabolism* ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Reactive Oxygen Species ; Rats ; Animals ; Myocytes, Cardiac/drug effects* ; Homocysteine/adverse effects* ; Hyperhomocysteinemia