Association between Age and Miscarriage in an Assisted Reproductive Technology Population: A 10-Year Cohort Study.

Background: Advanced maternal age decreased success of pregnancy rate in the assisted reproductive technology (ART) treatment. We aimed to investigate the association between age and miscarriages in women who required ART for conception. Methods: A cohort study was conducted using a 10-year sample of 14,898 pregnancy cycles with ART treatment in Xi'an, China. The effects of women's age on miscarriage were assessed using Poisson regression models. The threshold effect between age and miscarriage was explored through curve fitting. Results: Compared with lower than 30 years, the risks of early miscarriage and miscarriage were higher in the older age groups (early miscarriage: [35-37 years: RR=1.48, 95% confidence interval (CI): 1.26 to 1.74; ≥38 years: RR=2.25, 95% CI: 1.87 to 2.72]; miscarriage: [35-37 years: RR=1.45, 95%CI: 1.24 to 1.69; ≥38 years: RR=2.17, 95%CI: 1.82 to 2.60]). The nonlinear relationship between age and early miscarriage and miscarriages were observed. The risk of early miscarriage and miscarriage rapidly increased with age after the turning point (age=33 years) (<33 years: [early miscarriage: RR=1.02, 95% CI: 1.00 to 1.04; miscarriage: RR=1.02, 95% CI: 1.00 to 1.03]; ≥33 years: [early miscarriage: RR=1.11, 95% CI: 1.08 to 1.13; miscarriage: RR=1.10, 95% CI: 1.07 to 1.13]). Conclusion: Among pregnancy cycles undergoing ART, advanced age is associated with higher risk of early miscarriage and miscarriage, and nonlinear relationship between age and early miscarriage and miscarriage were found. More attention should be paid to the risk of pregnant women with older than 33 years in ART treatment.

The association between prepregnancy dietary fatty acids and risk of gestational diabetes mellitus: A prospective cohort study.

BACKGROUND & AIMS: Epidemiologic studies have examined the association between dietary fatty acids and type 2 diabetes risk in general populations. Evidence regarding their associations with gestational diabetes mellitus (GDM) risk remains limited. This study aimed to evaluate prepregnancy fatty acids intake in relation to GDM risk. METHODS: 3,725 pregnant women from the Xi'an Birth Cohort Study who were free of previous GDM or pre-existing chronic diseases were included. Dietary intake of total fat and individual fatty acids (including saturated fatty acids [SFA], monounsaturated fatty acids [MUFA], polyunsaturated fatty acids [PUFA], and trans fatty acids) during the year preceding pregnancy was assessed by a validated food-frequency questionnaire before 16 weeks of gestation. GDM was confirmed based on the 75-g oral glucose tolerance test. Log-binomial or modified Poisson regression models were applied to estimate the relative risks (RRs) and 95 % confidence intervals (95%CIs) of GDM for fatty acids intake. Generalized linear regression was adopted for blood glucose levels with fatty acids intake. RESULTS: 644 (17.3 %) incident GDM cases were confirmed in our study. Participants in the highest intake of total fat substituting for carbohydrates had a 33 % reduced risk of GDM than those in the lowest intake (RR:0.67; 95%CI:0.55,0.81). For individual fatty acids, only PUFA intake was associated with a lower risk of GDM, with RR comparing extreme tertiles of 0.61 (95%CI:0.49,0.76). Each 2 % increase in energy from total fat and PUFA replacing carbohydrates decreased the risk of GDM by 6 % (95%CI:3 %,9 %) and 15 % (95%CI:9 %,21 %), respectively. Similar inverse associations with intake of total fat and PUFA were observed for blood glucose levels. Further analyses of SFA substitution showed that replacement of 2 % energy from SFA with PUFA and MUFA was associated with 26 % (RR:0.74; 95%CI:0.62,0.88) and 30 % (RR:0.70; 95%CI:0.50, 0.98) decreased risk of GDM, respectively. CONCLUSIONS: Greater intake of total fat and PUFA before pregnancy was associated with lower risk of GDM when replacing carbohydrates. Substitution SFA with PUFA and MUFA was also inversely associated with GDM risk. These findings support the important role of optimal dietary fatty acids composition in the prevention of GDM.

Potassium levels and the risk of all-cause and cardiovascular mortality among patients with cardiovascular diseases: a meta-analysis of cohort studies.

BACKGROUND: Abnormal blood potassium levels are associated with an increased risk of cardiometabolic diseases and mortality in the general population; however, evidence regarding the association between dyskalemia and mortality among patients with cardiovascular disease (CVD) remains inconclusive. This study aimed to evaluate the association of potassium levels with all-cause and cardiovascular mortality among patients with CVD. METHODS: PubMed, Embase, Web of Science, and Cochrane Library databases were searched up to August 2023 to identify relevant cohort studies among patients with CVD, such as myocardial infarction, stroke, and heart failure. Abnormal potassium levels were considered as hypokalemia or hyperkalemia. The primary outcomes were all-cause mortality based on follow-up length (including in-hospital, short-term and long-term mortality) and cardiovascular mortality. The methodological quality of included studies was assessed by using the Newcastle-Ottawa Scale. The pooled relative risks (RRs) and 95% confidence intervals (CIs) were calculated using random-effects models. Restricted cubic splines were applied to explore the dose-response relationship. RESULTS: Thirty-one cohort studies involving 227,645 participants with an average age of 68.3 years were included in the meta-analysis, all of which achieved moderate to high quality. Hyperkalemia was significantly associated with an approximately 3.0-fold increased risk of all-cause in-hospital mortality (RR:2.78,95CI%:1.92,4.03), 1.8-fold of all-cause short-term mortality (RR:1.80, 95CI%:1.44,2.27), 1.3-fold of all-cause long-term mortality (RR:1.33, 95CI%:1.19,1.48) and 1.2-fold of cardiovascular mortality (RR:1.19, 95CI%:1.04,1.36). Similar positive associations were also observed between hypokalemia and risk of all-cause mortality and cardiovascular mortality. The RRs of all-cause in-hospital, short-term, long-term mortality and cardiovascular mortality with hyperkalemia were attenuated to 2.21 (95CI%:1.60,3.06), 1.46(95CI%:1.25,1.71), 1.23 (95CI%:1.09,1.39) and 1.13 (95CI%:1.00,1.27) when treating hypokalemia together with normokalemia as the reference group. A U-shaped association was observed between potassium levels and mortality, with the lowest risk at around 4.2 mmol/L. CONCLUSIONS: Both hypokalemia and hyperkalemia were positively associated with the risk of mortality in patients with CVD. Our results support the importance of potassium homeostasis for improving the CVD management. REGISTRATION: PROSPERO, CRD42022324337.

Potassium status and the risk of type 2 diabetes, cardiovascular diseases, and mortality: a meta-analysis of prospective observational studies.

Epidemiological evidence on the association between potassium and cardiometabolic outcomes remains controversial. This study aimed to examine associations of dietary intake and blood and urinary levels of potassium with risk of type 2 diabetes, cardiovascular disease (CVD), and mortality. Relevant prospective studies were retrieved through a comprehensive search of four electronic databases up to July 1, 2023. Random-effects models were used to pool the study-specific relative risks (RRs) and 95% confidence intervals (CIs). Fifty-six studies were included in this meta-analysis. A higher intake of potassium was significantly associated with a 16% lower risk of CVD (RR: 0.84, 95% CI: 0.78-0.90). Similar inverse associations were also observed between potassium intake and mortality. Each 1.0 g/d increment in potassium intake was associated with a decreased risk of CVD (RR: 0.85, 95% CI: 0.80-0.91) and all-cause mortality (RR: 0.93, 95% CI: 0.88-0.99). For blood and urinary potassium levels, higher level of blood potassium increased the risk of all-cause mortality by 23% (RR: 1.23, 95% CI: 1.11-1.36). The association of blood potassium levels with mortality was nonlinear (Pnon-linearit<0.001). However, urinary potassium levels were inversely associated with the risk of all-cause mortality (RR: 0.84, 95% CI: 0.76-0.93). Our findings support the benefits of moderate potassium consumption for primary prevention of chronic diseases and premature death.

Association of Plasma Vitamins and Carotenoids, DNA Methylation of LCAT, and Risk of Age-Related Macular Degeneration.

Dysregulation of lipid metabolism has been implicated in age-related macular degeneration (AMD), the leading cause of blindness among the elderly. Lecithin cholesterol acyltransferase (LCAT) is an important enzyme responsible for lipid metabolism, which could be regulated by DNA methylation during the development of various age-related diseases. This study aimed to assess the association between LCAT DNA methylation and the risk of AMD, and to examine whether plasma vitamin and carotenoid concentrations modified this association. A total of 126 cases of AMD and 174 controls were included in the present analysis. LCAT DNA methylation was detected by quantitative real-time methylation-1specific PCR (qMSP). Circulating vitamins and carotenoids were measured using reversed-phase high-performance liquid chromatography (RP-HPLC). DNA methylation of LCAT was significantly higher in patients with AMD than those in the control subjects. After multivariable adjustment, participants in the highest tertile of LCAT DNA methylation had a 5.37-fold higher risk (95% CI: 2.56, 11.28) of AMD compared with those in the lowest tertile. Each standard deviation (SD) increment of LCAT DNA methylation was associated with a 2.23-fold (95% CI: 1.58, 3.13) increased risk of AMD. There was a J-shaped association between LCAT DNA methylation and AMD risk (Pnon-linearity = 0.03). Higher concentrations of plasma retinol and ß-cryptoxanthin were significantly associated with decreased levels of LCAT DNA methylation, with the multivariate-adjusted ß coefficient being -0.05 (95% CI: -0.08, -0.01) and -0.25 (95% CI: -0.42, -0.08), respectively. In joint analyses of LCAT DNA methylation and plasma vitamin and carotenoid concentrations, the inverse association between increased LCAT DNA methylation and AMD risk was more pronounced among participants who had a lower concentration of plasma retinol and ß-cryptoxanthin. These findings highlight the importance of comprehensively assessing LCAT DNA methylation and increasing vitamin and carotenoid status for the prevention of AMD.

Consumption of sugar sweetened beverages, artificially sweetened beverages and fruit juices and risk of type 2 diabetes, hypertension, cardiovascular disease, and mortality: A meta-analysis.

Introduction: Sugar-sweetened beverage (SSB) intake is associated with an increased risk of cardiometabolic diseases. However, evidence regarding associations of artificially sweetened beverages (ASBs) and fruit juices with cardiometabolic diseases is mixed. In this study, we aimed to investigate the association between the SSB, ASB and fruit juice consumption with the incidence of cardiometabolic conditions and mortality. Methods: Relevant prospective studies were identified by searching PubMed, Web of Science, Embase, and Cochrane Library until December 2022 without language restrictions. The pooled relative risk (RR) and 95% confidence intervals (CIs) were estimated for the association of SSBs, ASBs, and fruit juices with the risk of type 2 diabetes (T2D), cardiovascular disease (CVD), and mortality by using random-effect models. Results: A total of 72 articles were included in this meta-analysis study. Significantly positive associations were observed between the consumption of individual beverages and T2D risk (RR: 1.27; 95% CI: 1.17, 1.38 for SSBs; RR: 1.32; 95% CI: 1.11, 1.56 for ASBs; and RR:0.98; 95% CI: 0.93, 1.03 for fruit juices). Moreover, our findings showed that intakes of SSBs and ASBs were significantly associated with risk of hypertension, stroke, and all-cause mortality (RR ranging from 1.08 to 1.54; all p < 0.05). A dose-response meta-analysis showed monotonic associations between SSB intake and hypertension, T2D, coronary heart disease (CHD), stroke and mortality, and the linear association was only significant between ASB consumption and hypertension risk. Higher SSB and ASB consumptions were associated with a greater risk of developing cardiometabolic diseases and mortality. Fruit juice intake was associated with a higher risk of T2D. Conclusion: Therefore, our findings suggest that neither ASBs nor fruit juices could be considered as healthier beverages alternative to SSBs for achieving improved health.Systematic Review Registration: [PROSPERO], identifier [No. CRD42022307003].

Prevalence of overweight, obesity, and associated factors among healthcare workers in the Gaza Strip, Palestine: A cross-sectional study.

Background: Overweight and obesity are multifactorial conditions that are prevalent in developing and developed countries. They are emerging as a significant public health concern among healthcare workers (HCWs). We aimed to estimate the prevalence of overweight and obesity and their associated factors among HCWs in the Gaza Strip. Methods: A cross-sectional study was conducted to recruit 1,850 HCWs aged 22 years and older. Interviews were carried out to collect sociodemographic information, nutritional information, and physical activity. Anthropometric measurements [height, weight, and waist circumference] were conducted with the HCWs. The body mass index was computed to determine the prevalence of overweight and obesity. Chi-square, t-test, and one-way ANOVA were used to compare the variables, and logistic regression was used to examine the associated factors of overweight and obesity. Results: The combined prevalence of overweight and obesity among HCWs was 65%. The result of logistic regression showed the risk of being overweight and obesity increased within the age group of 40-49 years (OR = 3.20; 95% CI: 2.37-4.32; P < 0.001). Male participants had more risk of obesity than female participants (OR = 1.77; 95% CI: 1.45-2.15). Married participants had a significantly higher risk of being overweight and obese (OR = 2.52; 95% CI: 2.05-3.28; P = 0.001). Increased monthly income was significantly associated with the risk of being overweight and obese (OR = 2.16; 95% CI: 1.22-3.83; P = 0.008). In addition, hypertension (OR = 2.49; 95% CI: 1.65-3.78; P < 0.001) and type 2 diabetes (OR = 2.42; 95% CI: 1.21-4.85; P= 0.012) were associated with overweight and obesity. Finally, a family history of NCDs was associated with overweight and obesity (OR = 1.69; 95% CI: 1.38-2.07; P < 0.001). Conclusion: This study showed a high prevalence of overweight and obesity among HCWs. Age, monthly income, marital status, known hypertension, type 2 diabetes, and eating habits were associated with the prevalence of overweight and obesity compared to other variables that were not associated with overweight and obesity such as profession, vegetables, fruit consumption, and physical activity. Urgent action is needed to tackle overweight and obesity among HCWs.

Dietary vitamins, carotenoids and their sources in relation to age-related macular degeneration risk in China: a population-based case-control study.

Mechanistic studies have suggested that antioxidants have beneficial effects on age-related macular degeneration (AMD). This study aimed to investigate the association between the types and sources of dietary vitamin and carotenoid intakes and AMD risk in China. A matched case-control study of 260 AMD cases and 260 matched controls was performed. The participants were interviewed for dietary information and potential confounders, and comprehensive ophthalmic examinations were performed. Conditional logistic models were used to estimate the odds ratio (OR) and 95 % confidence interval (CI) of specific vitamins and carotenoids and their main sources. When comparing the extreme quartiles, the ORs (95 % CI) were 0·30 (0·10, 0·88) for lutein and 0·28 (0·11, 0·74) for ß-cryptoxanthin. The associations for other dietary vitamin and carotenoid intakes were generally weaker and non-significant. Higher intakes of spinach and egg, which are important sources of lutein, were associated with a reduced odds of AMD. ORs (95% CIs) comparing extreme categories were 0·42 (0·20, 0·88) for spinach and 0·52 (95% CI: 0·27, 0·98) for egg. Participants who were in the highest category of both egg intake and spinach intake had a much greater reduced odds of having AMD (OR: 0·23; 95% CI: 0·08, 0·71) than those in the lowest category of egg intake and spinach intake. In conclusion, a higher intake of lutein and lutein-rich foods was associated with a significantly decreased odds of AMD. These findings provide further evidence of the benefits of lutein and lutein-rich foods in the prevention of AMD.

Dietary Habits of Patients with Coronary Artery Disease: A Case-Control Study from Pakistan.

BACKGROUND: Adults in South Asian countries have high chances of developing coronary artery disease (CAD) as compared to the developed nations. CAD is among the primary non-communicable causes of death in this region. Dyslipidemia, obesity, smoking hypertension, diabetes are considered as important risk factors for CVD. METHODS: A case-control study was conducted, with data was collected from the Punjab Institute of Cardiology in Lahore and the University of Lahore Teaching Hospital. A total of 500 subjects were selected, of which 250 were coronary artery disease patients and 250 were healthy controls. The CAD patients were selected from the outpatient department (OPD) and emergency unit of the Punjab Institute of Cardiology and the University of Lahore Teaching Hospital. RESULTS: The mean age of CAD patients was 57.83 ± 7.51 years and that of the controls was 55.32 ± 6.40 years. There was a significant difference in the mean values of biochemical parameters among cases and controls except for fasting blood sugar levels while there was a significant difference (p-value: 0.000) in the mean values of systolic blood pressure among cases and controls. Similarly, the values of diastolic blood pressure were also significantly different (p-value: 0.000) among cases and controls. The values of total blood cholesterol, LDL, triglycerides and HDL were also significantly different among cases and controls. There was a significant relationship between consumption of chicken, eggs, beef, yogurt, junk food, fresh vegetables, and fruits, and incidence of CAD. Consuming milk every day, and consuming fish weekly and consuming ghee had no significant association with the risk of coronary artery disease. On the other hand, from the findings of the unadjusted model, there was a significant association between CAD risk and intake of chicken, beef, egg, yogurt, junk food, fish, vegetables, and fruits. CONCLUSIONS: Diet is a risk factor for coronary artery disease and can be adjusted to reduce the risk of CAD. A key finding is that consumption of chicken, beef, eggs and junk food are associated with a high risk of CAD whereas consumption of ghee is not associated with the risk of CAD.

Intake of Soy, Soy Isoflavones and Soy Protein and Risk of Cancer Incidence and Mortality.

Background and Aims: Associations between soy intake and risk of cancer have been evaluated in prospective observational studies with inconsistent results. Whether the potential anticancer effects offered by soy were attributed to soy isoflavones and soy protein still needs to be elucidated. This study aimed to comprehensively quantify the association of soy, soy isoflavones and soy protein intake with risk of cancer incidence and cancer mortality by conducting a meta-analysis of all available studies. Methods: PubMed, Embase, Web of Science, and Cochrane Library databases were searched up to 16 September 2021. Prospective cohort studies that examined the effect of soy, soy isoflavones and soy protein on cancer incidence and cancer mortality were identified. Random-effects models were used to pool the multivariable-adjusted relative risks (RRs) and corresponding 95% confidence intervals (CIs). The potential dose-response relations were explored by using generalized least-squares trend estimation. Results: Eighty one prospective cohort studies were included in the meta-analysis. A higher intake of soy was significantly associated with a 10% reduced risk of cancer incidence (RR, 0.90; 95% CI, 0.83-0.96). Each additional 25 g/d soy intake decreased the risk of cancer incidence by 4%. Intake of soy isoflavones was inversely associated with risk of cancer incidence (RR, 0.94; 95% CI, 0.89-0.99), whereas no significant association was observed for soy protein. The risk of cancer incidence was reduced by 4% with each 10 mg/d increment of soy isoflavones intake. Similar inverse associations were also found for soy in relation to site-specific cancers, particularly lung cancer (RR, 0.67; 95%CI, 0.52-0.86) and prostate cancer (RR, 0.88; 95%CI, 0.78-0.99). However, high intake of soy, soy isoflavones and soy protein were not associated with cancer mortality. Conclusions: Higher intake of soy and soy isoflavones were inversely associated with risk of cancer incidence, which suggested that the beneficial role of soy against cancer might be primarily attributed to soy isoflavones. These findings support recommendations to include soy as part of a healthy dietary pattern for the prevention of cancer.

Prediction of occult tumor progression via platelet RNAs in a mouse melanoma model: a potential new platform for early detection of cancer.

BACKGROUND: Cancer screening provides the opportunity to detect cancer early, ideally before symptom onset and metastasis, and offers an increased opportunity for a better prognosis. The ideal biomarkers for cancer screening should discriminate individuals who have not developed invasive cancer yet but are destined to do so from healthy subjects. However, most cancers lack effective screening recommendations. Therefore, further studies on novel screening strategies are urgently required. METHODS: We used a simple suboptimal inoculation melanoma mouse model to obtain 'pre-diagnostic samples' of mice with macroscopic melanomas. High-throughput sequencing and bioinformatic analysis were employed to identify differentially expressed RNAs in platelet signatures of mice injected with a suboptimal number of melanoma cells (eDEGs) compared with mice with macroscopic melanomas and negative controls. Moreover, 36 genes selected from the eDEGs via bioinformatics analysis were verified in a mouse validation cohort via quantitative real-time PCR. LASSO regression was utilized to generate the prediction models with gene expression signatures as the best predictors for occult tumor progression in mice. RESULTS: These RNAs identified from eDEGs of mice injected with a suboptimal number of cancer cells were strongly enriched in pathways related to immune response and regulation. The prediction models generated by 36 gene qPCR verification data showed great diagnostic efficacy and predictive value in our murine validation cohort, and could discriminate mice with occult tumors from control group (area under curve (AUC) of 0.935 (training data) and 0.912 (testing data)) (gene signature including Cd19, Cdkn1a, S100a9, Tap1, and Tnfrsf1b) and also from macroscopic tumor group (AUC of 0.920 (training data) and 0.936 (testing data)) (gene signature including Ccr7, Cd4, Kmt2d, and Ly6e). CONCLUSIONS: Our proof-of-concept study provides evidence for potential clinical relevance of blood platelets as a platform for liquid biopsy-based early detection of cancer.

The Associations of Plasma Carotenoids and Vitamins With Risk of Age-Related Macular Degeneration: Results From a Matched Case-Control Study in China and Meta-Analysis.

BACKGROUND AND PURPOSE: Data from studies support a beneficial effect of carotenoids and vitamins on an age-related macular degeneration (AMD) risk. However, studies on the relations between blood levels of these nutrients and AMD are limited and provided conflicting results. The objective of this case-control study and meta-analysis was to examine whether the blood concentrations of carotenoids and vitamins were associated with the risk of AMD. METHODS: A total of 164 cases of AMD and an equal number of controls are individually matched according to age and gender among the participants, who provided blood samples in the Xi'an Eye Study. Plasma carotenoids and vitamins were measured using reversed-phase high-performance liquid chromatography. Bonferroni-corrected covariate-adjusted conditional logistic regression were used to estimate AMD risk by category of these nutrients in the multivariable-adjusted model. Nine studies were identified for the meta-analysis and calculated pooled risk estimates by means of a random-effects model. RESULTS: Plasma concentrations of examined carotenoids and vitamins were significantly lower in patients with AMD than those in controls. Plasma concentrations of examined carotenoids and vitamins were significantly lower in patients with AMD than those in controls. After a multivariate adjustment for body mass index, blood cholesterol, and other lifestyle risk factors, higher lutein/zeaxanthin content in plasma was significantly associated with a decreased risk of AMD, and the odds ratio (OR) comparing the top and bottom tertiles was 0.21 (95% CI: 0.05, 0.84; P trend = 0.024). Associations for ß-carotenes (OR: 0.11; 95% CI: 0.02, 0.50; P trend < 0.001), and ß-cryptoxanthin (OR: 0.08, 95% CI: 0.02, 0.39; P trend < 0.001) were similar to that for lutein/zeaxanthin. Inverse associations were also observed for a higher level of retinol (OR: 0.14, 95% CI: 0.03, 0.61; P trend = 0.006) and α-tocopherol (OR: 0.25, 95% CI: 0.06, 0.98; P trend = 0.006). In the meta-analysis, a protective effect was detected for AMD among the participants with high blood lutein/zeaxanthin level (OR: 0.53, 95% CI: 0.40, 0.72, P < 0.001), compared to those with low level. Similar inverse associations were seen for ß-carotene (OR: 0.48, 95% CI: 0.28, 0.84, P = 0.010), ß-cryptoxanthin (OR: 0.48, 95% CI: 0.23, 1, P = 0.049), lycopene (OR: 0.70, 95% CI: 0.54, 0.90, P = 0.006) and α-tocopherol (OR: 0.50, 95% CI: 0.31, 0.81, P = 0.005). CONCLUSIONS: Results of the case-control study were consistent with findings from the meta-analysis, indicating that higher concentrations of carotenoids and vitamins were inversely associated with the AMD risk. Our finding supports the current notion that these nutrients are likely to affect the development of AMD and may help to refine the strategies for the prevention of age-related eye diseases.

An eQTL variant of ALDH1A2 is associated with Kashin-Beck disease in Chinese population.

INTRODUCTION: The aims of the study were to investigate the relationship between aldehyde dehydrogenase 1 family member A2 (ALDH1A2) and Kashin-Beck disease (KBD), explore the effects of the rs3204689 polymorphism and methylation status on the expression levels of ALDH1A2, and further clarify the pathogenesis of KBD. MATERIALS AND METHODS: The genotype of ALDH1A2 rs3204689 was detected by PCR-RFLP in 103 KBD patients and 109 healthy controls in the whole blood. The mRNA level of ALDH1A2 was measured by qRT-PCR, and the protein expression was detected using IHC staining and Western blotting. The MSP-PCR was used to identify the ALDH1A2 methylation level. RESULTS: There were significant differences in G/G, G/C, and C/C frequencies of ALDH1A2 rs3204689 between the KBD and control groups (χ2 = 7.113, P = 0.029); the minor allele G of ALDH1A2 was associated with the risk of KBD (χ2 = 5.984, P = 0.014). The mRNA and protein levels of ALDH1A2 were increased in the whole blood and cartilage of KBD patients compared with the controls (P = 0.049, P < 0.0001, P = 0.019). Meanwhile, a statistically significant difference was observed between G/G, G/C and C/C genotype on mRNA expression (P = 0.039). The methylation level of the ALDH1A2 gene promoter region showed no significant difference between the KBD and control groups (χ2 = 0.317, P = 0.573). CONCLUSION: Our case-control study indicates that the common variant rs3204689 near ALDH1A2 is associated with KBD in Chinese population. The risk allele G of rs3204689 is statistically linked to the high expression of ALDH1A2, which is up-regulated in the cartilage and whole blood of KBD patients. Our findings suggest a potential role of ALDH1A2 in the pathogenesis of KBD.

Dysregulation of Transcription Profile of Selenoprotein in Patients with Kashin-Beck Disease and Its Effect on Se Deficiency-Induced Chondrocyte Apoptosis.

Kashin-Beck disease (KBD) is a chronic, degenerative osteoarthropathy related to selenium (Se) deficiency. Se participates in the synthesis of selenoprotein in the form of selenocysteine. In total, 25 selenoproteins, encoded by 25 genes, are currently found in humans; however, the effects of selenoprotein genes on chondrocyte apoptosis, particularly in apoptosis-related genes, remain poorly elucidated. Therefore, in the current study, the expression of selenoprotein genes and apoptosis-related genes were determined by RT-qPCR in patients and chondrocytes and the correlations between them were analyzed using Pearson and Spearman's rank correlation, and the chondrocyte apoptosis rate was detected by Annexin V-FITC/PI. The results showed that the mRNA levels of 17 selenoprotein genes were downregulated, whereas two genes were upregulated in patients with KBD. The BAX/BCL2 ratio and the mRNA levels of BAX and P53 were increased, but the mRNA levels of BCL2 and NF-κB p65 were decreased in patients with KBD. The mRNA levels of GPX2, GPX3, DIO1, TXNRD1, TXNRD3, and SPS2 were most closely associated with apoptosis-related genes in patients with KBD. Moreover, in the Se deficiency group, the mRNA levels of GPX3, DIO1, and TXNRD1 were downregulated and GPX activity was decreased, but the late apoptosis rate, the mRNA levels of BAX and P53, and the BAX/BCL2 ratio were increased; the opposite trend was observed in the Se supplement group. Collectively, these results indicate that selenoprotein transcription profile is dysregulated in patients with KBD. Furthermore, the expression of GPX3, DIO1, and TXNRD1 genes might be involved in the development of chondrocyte apoptosis by affecting antioxidant capacity.

Identification of differentially expressed and methylated genes associated with rheumatoid arthritis based on network.

Rheumatoid arthritis (RA) is a multi-systemic inflammatory autoimmune disease involving peripheral joints, and the pathogenesis is not clear. Studies showed that DNA methylation and expression might also be involved in the pathogenesis of RA. This study integrated three expression profile datasets (GSE55235, GSE12021, and GSE55457) and one methylation profile dataset GSE111942 to elucidate the potential essential candidate genes and pathways in RA. Differentially expressed genes (DEGs) and differentially methylation genes (DMGs) were identified by R programming software, using Limma package and ChAMP package, respectively. DAVID performed gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs. Functional annotation and construction of a protein-protein interaction (PPI) network and the Molecular Complex Detection Algorithm (MCODE) were analysed by STRING and Cystoscope, respectively. Then the connection analysis of DEGs and DMGs was carried out, and further to analyse the relationship between methylation and gene expression, aiming to screen out the potential genes. In this study, 288 DEGs and 228 DMGs were identified, and the majority of DEGs were up-regulated. Enrichment analysis represented that DEGs mainly involved immune response and participated in the Cytokine-cytokine receptor interaction signal pathway. 282 nodes were identified from DEGs PPI network and MCODE, filtering the most significant 2 modules, 23 core node genes were identified and most of them are involved in the T cell receptor signalling pathway and chemokine-mediated signalling pathway. Cross-analysis revealed 4 genes [KNTC1 (cg 01277763), LRRC8D (cg 07600884), DHRS9 (cg 05961700), and UCP2 (cg 05205664)] that exhibited differential expression and methylation in RA simultaneously. Therefore, the four genes could be used as the target for RA.

Effects of methylation of deiodinase 3 gene on gene expression and severity of Kashin-Beck disease.

Kashin-Beck disease (KBD) is a complex endemic osteoarthropathy, which mainly occurs in the northeast to southwest China. Iodothyronine deiodinases 3 (DIO3) is one of the selenoproteins, which is closely related to bone metabolism and unclear to KBD. This study aims to investigate the role and associated mechanisms of methylation and expression of DIO3 with disease severity in patients with KBD. We performed a bioinformatics analysis first to identify the biological mechanisms involved in selenoproteins. The methylation status of the DIO3 gene and DIO3 gene expression, as well as DIO3-related regulatory genes in patients with KBD, were analyzed. We found that 15 CpG sites of six selenoproteins were hypomethylated with 5-azacytidine treatment. DIO3 hypermethylation was associated with an increased risk of KBD and may lead to downregulation of DIO3 gene expression as well as be an indicator of the severity of KBD, which may provide a new insight for gene-environment correlations and interactions in etiology and pathogenesis of KBD.

No Association between Estrogen Receptor-Β Rs4986938 and Cancer Risk: A Systematic Review and Meta-Analysis.

BACKGROUND: The association between estrogen receptor-ß (ESR2) rs4986938 polymorphism and the risk of various types of cancer have been investigated in previous studies. However, the results remained disputable. Here, we conducted a meta-analysis to investigate the association between ESR2 rs4986938 polymorphism and the risk of cancer. METHODS: We searched for relevant articles collected by the PubMed, EMBASE, and Cochrane library up to March 30, 2018. The association was assessed using Odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: The meta-analysis involved a total of 23 studies in 20 papers, including 24,334 cases and 31,707 controls. No significant association was detected between the rs4986938 polymorphism and cancer risk in the additive model (A compared with G: OR=0.97, 95% CI=0.92-1.02, P=0.20), dominant model (AA+AG compared with GG: OR=0.96, 95% CI=0.93-1.03, P=1.00), recessive model (AA compared with AG + GG: OR=0.94, 95% CI=0.86-1.03, P=0.18), heterozygous model (AG compared with GG: OR=0.97, 95% CI=0.94-1.01, P=0.14), and homozygous model (AA compared with GG: OR=0.96, 95% CI=0.87-1.06, P=0.39). Results of subgroup analysis stratified by ethnicity and cancer types further validated the results. CONCLUSION: We found no evidence of an association between rs4986938 and the risk of overall cancer.

Pathway-based network analyses and candidate genes associated with Kashin-Beck disease.

To perform a comprehensive analysis focusing on the biological functions and interactions of Kashin-Beck disease (KBD)-related genes to provide information towards understanding the pathogenesis of KBD.A retrospective, integrated bioinformatics analysis was designed and conducted. First, by reviewing the literature deposited in PubMed, we identified 922 genes genetically associated with KBD. Then, biological function and network analyses were conducted with Cytoscape software. Moreover, KBD specific molecular network analysis was conducted by Cytocluster using the Molecular Complex Detection Algorithm (MCODE).The biological function enrichment analysis suggested that collagen catabolic process, protein activation cascade, cellular response to growth factor stimulus, skeletal system development, and extrinsic apoptosis played important roles in KBD development. The apoptosis pathway, NF-kappa B signaling pathway, and the glutathione metabolism pathway were significantly enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway network, suggesting that these pathways may play key roles in KBD occurrence and development. MCODE clusters showed that in top 3 clusters, 54 of KBD-related genes were included in the network and 110 candidate genes were discovered might be potentially related to KBD.The 110 candidate genes discovered in the current study may be related to the development of KBD. The expression changes of apoptosis and oxidative stress-related genes might serve as biomarkers for early diagnosis and treatment of KBD.

Potential candidate biomarkers associated with osteoarthritis: Evidence from a comprehensive network and pathway analysis.

Osteoarthritis (OA) is one of the most common forms of arthritis world widely. Some key genes and diagnostic markers have been reported due to the development of modern molecular biology technologies. However, the etiology and pathogenesis of OA remains unknown. In this study, an integrated network and pathway analysis towards the biological function of OA-associated genes was conducted to provide valuable information to further explore the etiology and pathogenesis of OA. A total of 2,548 genes which reported a statistically significant association with OA were screened. An integrated network and pathway analysis was performed to identify the pathways and genes most associated to OA. Moreover, OA-specific protein-protein interaction (PPI) network was constructed by cytocluster based on the Molecular Complex Detection Algorithm (MCODE) to screen its candidate biomarkers. Quantitative real-time polymerase chain reaction was used to confirm the expression levels and to validate the results of MCODE cluster analysis by six genes. The pathway networks suggested that extracellular matrix (ECM) organization, collagen degradation and collagen formation showed important associations with OA. In top two PPI clusters, 61 of the OA-associated genes were included in the OA-specific PPI network, which also included 23 candidate genes that are likely to be highly associated with OA based on MCODE clusters. Analysis of mRNA showed that the expression levels of COL9A1, COL9A2, ITGA3, COL9A3, ITGA2, and LAMA1 in the peripheral blood mononuclear cells of OA patients were significantly lower than those of the normal controls (p<0.005). To our knowledge, this is the first comprehensive and systematic report based on OA-related genes demonstrating that the functional destruction of collagen in cartilage may be a very important contributing factor to OA. Quantitative detection of collagen synthesis may be of great help in early identification and prediction of OA. Maintaining the quality and quantity of collagen can be a potential target for clinical treatment of OA in the future practice.

Analysis of gene expression and methylation datasets identified ADAMTS9, FKBP5, and PFKBF3 as biomarkers for osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a kind of chronic osteoarthropathy and degenerative joint disease. Epigenetic regulation in the gene expression dynamics has become increasingly important in OA. We performed a combined analysis of two types of microarray datasets (gene expression and DNA methylation) to identify methylation-based key biomarkers to provide a better understanding of molecular biological mechanisms of OA. METHODS: We obtained two expression profiling datasets (GSE55235, GSE55457) and one DNA methylation profiling data set (GSE63695) from the Gene Expression Omnibus. First, differentially expressed genes (DEGs) between patients with OA and controls were identified using the Limma package in R(v3.4.4). Then, function enrichment analysis of DEGs was performed using a DAVID database. For DNA methylation datasets, ChAMP methylation analysis package was used to identify differential methylation genes (DMGs). Finally, a comprehensive analysis of DEGs and DMGs was conducted to identify genes that exhibited differential expression and methylation simultaneously. RESULTS: We identified 112 DEGs and 2,896 DMGs in patients with OA compared with controls. Functional analysis of DEGs obtained that inflammatory responses, immune responses, and positive regulation of apoptosis, tumor necrosis factor (TNF) signaling pathway, and osteoclast differentiation may be involved in the pathogenesis of OA. Cross-analysis revealed 26 genes that exhibited differential expression and methylation in OA. Among them, ADAMTS9, FKBP5, and PFKBF3 were identified as valuable methylation-based biomarkers for OA. CONCLUSION: In summary, our study identified different molecular features between patients with OA and controls. This may provide new clues for clarifying the pathogenetic mechanisms of OA.