The Epidemic Characteristics of Stroke Death from 2012 to 2021 in Chongqing, China.

INTRODUCTION: Stroke has become a major disease that threatens the global population's health and is a major public health problem that needs to be solved in China. Therefore, it is essential to analyze the trend of the mortality of stroke and its epidemic characteristic of stroke death. METHODS: Death cases of stroke were reported to the national death registry system by the medical staff of all medical institutions, and the population data every year were obtained from District or County's Statistic Bureau in Chongqing. They were analyzed to calculate the mortality, age-standardized mortality rate by Chinese standardization population (ASMRC), age-specific mortality, proportion, and annual percent of change (APC) according to the ICD-10 code. ASMRC was based on the standard population of the 6th census in China, 2010. The stroke mortality of each subgroup was compared using the χ2 test. Trend analysis was presented by APC. RESULTS: The crude mortality of stroke increased from 96.29 per 100,000 in 2012 to 115.93 per 100,000 significantly, with the APC of 2.02% (t = 2.82, p = 0.022) in Chongqing. ASMRC of stroke was 56.47 per 100,000 in 2012 and 54.70 per 100,000 in 2021, and its trend change was stable (APC = -0.01, t = 0.07, p = 0.947). The crude mortality of stroke in males was higher than that in females every year (p < 0.05). The death proportion of intracerebral hemorrhage dwindled from 60.53% in 2012 to 49.88% in 2021, whereas the death proportion of ischemic stroke increased from 20.92% in 2012 to 39.96% in 2021. The average age of stroke death was delayed from 73.43 years old in 2012 to 76.52 years old in 2021 significantly (t = 18.12, p < 0.001). The percentage of stroke death at home increased from 75.23% in 2012 to 79.23% in 2021, while the percentage of stroke death at hospitals decreased from 17.89% in 2012 to 15.89% in 2021. CONCLUSION: The crude mortality of stroke surged, and intracerebral hemorrhage was the main death cause of all subtypes. The mortality of stroke in males and rural residents was higher than that in females and urban residents. Most stroke deaths occurred at home. Male and rural residents were crucial populations for stroke prevention and control. There should be improved medical resources in rural areas and enhanced capability of stroke diagnosis and treatment.

Associations between major dietary patterns and blood pressure among Southwest Chinese: A cross-sectional analysis based on the China Multi-Ethnic Cohort (CMEC) study.

BACKGROUND AND AIMS: In the less developed multi-ethnic regions (LEMRs) of Southwest China, the associations between dietary patterns and blood pressure (BP) values remain unclear. We aimed to investigate such associations and related effect modifiers. METHODS AND RESULTS: This study included 81,433 participants from the China Multi-Ethnic Cohort Study. Dietary intakes during the year before the interview were measured with the Quantitative Food Frequency Questionnaire. Three major dietary patterns that were highly in line with geographical and ethnic distributions of the study population, i.e., "Sichuan Basin," "Yunnan-Guizhou Plateau," and "Qinghai-Tibet Plateau," were derived using principal component factor analysis. The multilinear regression model combined with inverse probability of exposure weighting was used to estimate the associations between dietary patterns and BP values. Comparing the highest with the lowest quintiles, the Sichuan Basin dietary pattern (characterized by urban lifestyles) was associated with 2.67 mmHg lower systolic blood pressure (SBP) (95% CI: -3.07 to -2.27) and 0.89 mmHg lower diastolic blood pressure (95% CI: -1.12 to -0.65). In contrast, both the Yunnan-Guizhou Plateau (characterized by agricultural lifestyles) and the Qinghai-Tibet Plateau dietary patterns (characterized by nomadic lifestyles) showed positive associations with BP. In the stratified analysis, the associations between dietary patterns and SBP were significantly stronger in women than in men for all three kinds of dietary patterns. CONCLUSION: Both major dietary patterns and their associations with BP showed a substantial disparity in LEMRs of Southwest China. Dietary patterns in regions of higher socioeconomic status are more conducive to reducing the rising of BP, especially for women and urban residents, which might provide insights into the BP control in LEMRs of Southwest China.

[Ox-LDL/ß2GPI/anti-ß2GPI antibody complex promotes apoptosis and inflammatory factors expression in RAW264.7 cells].

Objective To investigate the effects of ox-LDL/ß2GPI/anti-ß2GPI antibody complex on apoptosis and inflammatory factors expression in RAW264.7 cells and its mechanism. Methods RAW264.7 cells were treated with medium, ox-LDL, ß2GPI/aß2GPI antibody complex, ox-LDL/ß2GPI complex, ox-LDL/aß2GPI antibody complex, and ox-LDL/ß2GPI/aß2GPI antibody complex separately. The role of signaling pathways was investigated by TLR4 inhibitor TAK-242 and mTOR inhibitor rapamycin. Finally, cells were stimulated with different concentrations of TNF-α. The apoptosis was detected by Annexin V-FITC/PI double labeling. The mRNA expressions of TNF-α and IL-1ß were detected by real-time quantitative PCR, and the protein expressions of TNF-α, IL-1ß, and apoptosis-related proteins cleaved-caspase-3 (c-caspase-3) and Bcl2 were detected by Western blot. Results Ox-LDL/ß2GPI/aß2GPI antibody complex promoted RAW264.7 cells' apoptosis, with a decrease in protein Bcl2 and an increase in protein cleaved-caspase-3, and enhanced the level of inflammatory factors, while apoptosis and the expression of inflammatory factors were partially reduced when macrophages had been pre-treated with inhibitors. TNF-α up-regulated apoptosis in RAW264.7 cells at 100 ng/mL and 200 ng/mL. Conclusion Ox-LDL/ß2GPI/aß2GPI antibody complex induces apoptosis in RAW264.7 cells by increasing the expression of inflammatory factors. TLR4 and mTOR pathways are involved in the process.

Impaired Autophagy Induced by oxLDL/ß2GPI/anti-ß2GPI Complex through PI3K/AKT/mTOR and eNOS Signaling Pathways Contributes to Endothelial Cell Dysfunction.

Endothelial cell dysfunction plays a fundamental role in the pathogenesis of atherosclerosis (AS), and endothelial autophagy has protective effects on the development of AS. Our previous study had shown that oxidized low-density lipoprotein/ß2-glycoprotein I/anti-ß2-glycoprotein I antibody (oxLDL/ß2GPI/anti-ß2GPI) complex could promote the expressions of inflammatory cytokines and enhance the adhesion of leukocytes to endothelial cells. In the present study, we aimed to assess the effects of oxLDL/ß2GPI/anti-ß2GPI complex on endothelial autophagy and explore the associated potential mechanisms. Human umbilical vein endothelial cells (HUVECs) and mouse brain endothelial cell line (bEnd.3) were used as models of the vascular endothelial cells. Autophagy was evaluated by examining the expressions of autophagic proteins using western blotting analysis, autophagosome accumulation using transmission electron microscopy, and RFP-GFP-LC3 adenoviral transfection and autophagic flux using lysosome inhibitor chloroquine. The expressions of phospho-PI3K, phospho-AKT, phospho-mTOR, and phospho-eNOS were determined by western blotting analysis. 3-Methyladenine (3-MA) and rapamycin were used to determine the role of autophagy in oxLDL/ß2GPI/anti-ß2GPI complex-induced endothelial cell dysfunction. We showed that oxLDL/ß2GPI/anti-ß2GPI complex suppressed the autophagy, evidenced by an increase in p62 protein, a decrease in LC3-II and Beclin1, and a reduction of autophagosome generation in endothelial cells. Moreover, inhibition of autophagy was associated with PI3K/AKT/mTOR and eNOS signaling pathways. Rapamycin attenuated oxLDL/ß2GPI/anti-ß2GPI complex-induced endothelial inflammation, oxidative stress, and apoptosis, whereas 3-MA alone induced the endothelial injury. Our results suggested that oxLDL/ß2GPI/anti-ß2GPI complex inhibited endothelial autophagy via PI3K/AKT/mTOR and eNOS signaling pathways and further contributed to endothelial cell dysfunction. Collectively, our findings provided a novel mechanism for vascular endothelial injury in AS patients with an antiphospholipid syndrome (APS) background.

OxLDL/ß2GPI/anti­ß2GPI Ab complex induces inflammatory activation via the TLR4/NF­κB pathway in HUVECs.

Patients with antiphospholipid syndrome have been identified to have higher incidence rates of atherosclerosis (AS) due to the elevated levels of anti­ß2­glycoprotein I (ß2GPI) antibody (Ab). Our previous studies revealed that the anti­ß2GPI Ab formed a stable oxidized low­density lipoprotein (oxLDL)/ß2GPI/anti­ß2GPI Ab complex, which accelerated AS development by promoting the accumulation of lipids in macrophages and vascular smooth muscle cell. However, the effects of the complex on endothelial cells, which drive the initiation and development of AS, remain unknown. Thus, the present study aimed to determine the proinflammatory roles of the oxLDL/ß2GPI/anti­ß2GPI Ab complex in human umbilical vein endothelial cells (HUVECs) in an attempt to determine the underlying mechanism. Reverse transcription­quantitative PCR, enzymy­linked immunosorbent assay, western blotting and immunofluorescence staining were performed to detect the expressions of inflammation related factors and adhesion molecules. Monocyte­binding assay was used to investigate the effects of oxLDL/ß2GPI/anti­ß2GPI Ab complex on monocyte adhesion to endothelial cells. The results demonstrated that the oxLDL/ß2GPI/anti­ß2GPI Ab complex upregulated the expression of Toll­like receptor (TLR)4 and the levels of NF­κB phosphorylation in HUVECs, and subsequently enhanced the expression levels of inflammatory cytokines, including TNF­α, IL­1ß and IL­6, as well as those of adhesion molecules, such as intercellular adhesion molecule 1 and vascular adhesion molecule 1. In addition, the complex facilitated the recruitment of monocytes by promoting the secretion of monocyte chemotactic protein 1 in HUVECs. Notably, the described effects of the oxLDL/ß2GPI/anti­ß2GPI Ab complex in HUVECs were abolished by either TLR4 or NF­κB blockade. In conclusion, these findings suggested that the oxLDL/ß2GPI/anti­ß2GPI Ab complex may induce a hyper­inflammatory state in endothelial cells by promoting the secretion of proinflammatory cytokines and monocyte recruitment, which was discovered to be largely dependent on the TLR4/NK­κB signaling pathway.

Excessive gestational weight gain in the first and second trimester is a risk factor for gestational diabetes mellitus among women pregnant with singletons: A repeated measures analysis.

AIMS/INTRODUCTION: To evaluate gestational weight gain (GWG) in the first and second trimester as a risk factor for gestational diabetes mellitus (GDM) among women pregnant with singletons. MATERIALS AND METHODS: This was a cohort study of women with singleton pregnancies who delivered between 1 January 2013 and 31 October 2014 in a Chinese hospital. We collected data from medical records from the first antenatal visit to delivery. All pregnant women were subjected to an oral glucose tolerance test for diagnosis of GDM during the second trimester. GWG in the first and second trimester was calculated by subtracting the prepregnancy weight from weight within 4 weeks of the oral glucose tolerance test. We categorized GWG into insufficient, appropriate and excessive according to the Institute of Medicine guidelines and population quantiles. Univariable and multivariable analyses were used to determine the association between GWG and GDM risk. RESULTS: Of 10,422 pregnant women, we identified 8,356 eligible women with 1,622 (19.4%) diagnosed with GDM. Univariable analysis showed that GWG that exceeded the Institute of Medicine recommendation might be associated with risk of GDM (P < 0.05), but this association was not observed by multivariable analysis (adjusted odds ratio 1.07, [95% confidence interval 0.94-1.21]). Univariable and multivariable analyses both showed that GWG exceeding the 90th and 95th quantiles of included women, respectively, were at increased risk for GDM (adjusted odds ratio >P90 vs P10 -P90 adjusted odds ratio 1.31, [95% confidence interval 1.12-1.52]; >P95 vs P5 -P95 adjusted odds ratio 1.45 [95% confidence interval 1.16-1.81]). CONCLUSIONS: Excessive GWG in the first and second trimester might be a risk factor for GDM, which highlights the importance of appropriate weight gain during pregnancy.

Perfluorooctanoic acid exposure impact a trade-off between self-maintenance and reproduction in lizards (Eremias argus) in a gender-dependent manner.

The trade-off between self-maintenance and reproduction has been explored wildly in reptiles. However, the effects of exogenous pollutants on the life history traits of reptiles have not been paid attention to. In the current study, lizards (Eremias argus), living in the soil polluted by perfluorooctanoic acid (PFOA) were selected as the main focus. Bodyweight, survival rate, clutch characteristics and biochemical analysis (immune response, lipid accumulation, sex steroid secretion, antioxidant level, and metabolomics) were investigated and the results revealed that lizards' life-history trade-offs are gender-dependent: females were more inclined to choose a "Conservative" life-history strategy. After 60 days of exposure to PFOA, larger body weight, higher survival rate, stronger immune response, and lighter egg mass in females suggested that their trade-offs are more biased towards self-maintenance. Whereas, the "Risk" strategy would more popular among males: reduced body weight and survival rate, and suffering from oxidative damage indicated that males made little investment in self-maintenance.

Effects of ß2/aß2 on oxLDL-induced CD36 activation in THP-1 macrophages.

AIMS: ß2-glycoprotein I/anti-ß2-glycoprotein I antibody complex (ß2/aß2) could promote oxLDL-induced endothelial inflammation through Toll-like receptor 4 (TLR4), therefore accelerates atherosclerosis in patients with anti-phospholipid syndrome (APS). However, effects of ß2/aß2 and TLR4 on oxLDL-induced CD36 activation in macrophages remain to be elucidated and are currently under investigation. MATERIALS AND METHODS: THP-1 macrophages with or without the pre-treatment of TAK-242, a TLR4 inhibitor, were treated with RPMI 1640, oxLDL, oxLDL+ß2/aß2 or oxLDL + LPS.CD36 expression and subsequent intracellular lipid accumulation, cholesterol-transportation-related proteins (ACAT1, ABCG1 and ABCA1) expression, inflammatory cytokines (IL-1ß, TNF-α and IL-6) secretion, focal adhesion kinases (FAK) activation and matrix metalloproteinases (MMP-2 and MMP-9) expression by these THP-1 macrophages were evaluated. Moreover, effects of TLR4 on oxLDL+ß2/aß2-induced peroxisome proliferators-activated receptor-γ (PPAR-γ) expression and CD36 translocation have also been observed. KEY FINDINGS: Compared with oxLDL-treated ones, CD36 expression, intracellular lipid accumulation and FAK activation were inhibited, whereas the levels of inflammatory cytokines and MMPs were upregulated in THP-1 macrophages treated with oxLDL+ß2/aß2 (p < 0.05). Moreover, observed differences between oxLDL-treated and oxLDL+ß2/aß2-treated THP-1 macrophages could be reversed by TAK-242 pre-treatment (p < 0.05). Furthermore, oxLDL+ß2/aß2 promoted PPAR-γ expression and CD36 cytoplasmic translocation in THP-1 macrophages, these effects could also be attenuated by TAK-242 (p < 0.05). SIGNIFICANCE: Through a TLR4 dependent manner, ß2/aß2 inhibited oxLDL-induced CD36 expression, lipid accumulation and FAK activation, while promoted inflammatory cytokines and MMPs expression in THP-1 macrophages, indicating the novel dual roles played by ß2/aß2 in APS-related atherosclerosis.

The use of rigorous methods was strongly warranted among prognostic prediction models for obstetric care.

OBJECTIVE: The objective of the study was to examine methodological characteristics about the design and conduct in prognostic prediction models used for obstetric care. STUDY DESIGN AND SETTING: We searched PubMed for studies on prognostic prediction models for obstetric care, published in top general medicine or major specialty journals between January 2011 and February 2018. Teams of method-trained investigators independently screened titles and abstracts and collected data using a prespecified, pilot-tested, structured questionnaire. RESULTS: In total, 91 studies were eligible, of which two were published in top general medicine journals, 20 (22.0%) involved an epidemiologist or statistician, 18 (19.4%) published study protocols, 53 (58.2%) did not include any model validation, 20 (22.0%) did not clearly state the intended timing of use, 23 (25.3%) had no eligibility criteria, 15 (16.5%) did not use clear criteria for ascertaining outcome, and 69 (75.82%) did not apply blinding to outcome assessment. Among those models, 11 (12.1%) included participants fewer than 200 events, 41 (48.8%) had fewer than 100 events, and 19 (24.7%) had fewer than 10 events per variable. CONCLUSION: The prognostic prediction models have important limitations in design and conduct. Substantial efforts are needed to strengthen the production of reliable prognostic prediction models for obstetric care.

The mediation effect of multiple gestations on the association between in vitro fertilisation and severe maternal morbidities: a retrospective cohort study.

OBJECTIVE: To investigate whether the association between in vitro fertilisation (IVF) and severe maternal morbidity (SMM) was mediated by multiple gestations. DESIGN: A retrospective cohort study. SETTING: The study was conducted at six hospitals in China. PARTICIPANTS: Pregnant women at 20 gestational weeks or longer. OUTCOME MEASURE: The outcome was SMM, which was a composite of potential life-threatening conditions, the use of critical medical interventions, or the status of maternal near-miss that occurred during pregnancy, childbirth or within 42 days of pregnancy termination, as defined by WHO. RESULTS: In total, 22 368 eligible pregnant women were included, among whom 497 (2.2%) received IVF, and 776 developed SMM (incidence 34.7/1000 live births, 95% CI 32.3/1000 to 37.1/1000). Four multivariable logistic regression models were constructed. Model 1, without including the variable of multiple gestations, showed that IVF was associated with higher risk of SMM (adjusted OR (aOR) 1.54, 95% CI 1.03 to 2.29). Model 2, assessing the association between IVF and multiple gestations, showed that IVF was strongly associated with multiple gestations (aOR 14.75, 95% CI 11.38 to 19.10). Model 3, by adding the variable of multiple gestations to model 1, showed that IVF was not statistically associated with SMM (aOR 0.89, 95% CI 0.58 to 1.36), but multiple gestations were associated with higher risk of SMM (aOR 5.92, 95% CI 4.88 to 7.83). Model 4, investigating the association between IVF and SMM among singleton pregnancies, showed no statistically significant association (aOR 0.70, 95% CI 0.37 to 1.32). An additional analysis by adding the interaction term of IVF by multiple gestations to model 3 showed no statistical significance of the interaction term (aOR 1.15, 95% CI 0.36 to 3.68), confirming the absence of exposure-mediator interaction. CONCLUSIONS: Using the established rule for judging mediation effect, the results suggested that multiple gestations might mediate the association between the use of IVF and higher risk of SMM. Further prospective studies are warranted to test our finding.

[The oxLDL/ß2GPI/anti-ß2GPI antibody complex induces apoptosis of human umbilical vein endothelial cells by promoting the production of reactive oxygen species].

Objective To investigate the effect of oxidative low-density lipoprotein/ß2 glycoprotein I/anti-ß2 glycoprotein I antibody (oxLDL/ß2GPI/αß2GPIAb) complex on the apoptosis of human umbilical vein endothelial cells (HUVECs) and the role of reactive oxygen species (ROS) in the process. Methods HUVECs were stimulated by ß2GPI, aß2GPIAb, ß2GPI/aß2GPIAb complex, oxLDL, oxLDL/ß2GPI complex, oxLDL/ß2GPI/rabbit (R)-IgG complex, and oxLDL/ß2GPI/aß2GPIAb complex. The protein levels of apoptosis-related proteins including BAX, Bcl2 and cleaved caspase-3 (c-caspase-3) were detected by Western blot analysis. Annexin V-FITC/PI double labeling combined with flow cytometry was performed to detect apoptosis rate; dichlorofluorescein diacetate (DCFH-DA) labeling combined with flow cytometry was used to detect ROS level. And the effect of each treatment on cell viability was analyzed by CCK-8 assay. The changes of mean fluorescence intensity of ROS and the apoptosis of HUVECs induced by oxLDL/ß2GPI/aß2GPIAb complex were observed by the pretreatment with antioxidant apocynin or diphenylphosphonium iodide (DPI). Results The oxLDL/ß2GPI/aß2GPIAb complex significantly reduced the viability of HUVECs, increased the expression of BAX and cleaved caspase-3 protein, decreased the level of Bcl2 protein, and promoted the apoptosis of HUVECs. In addition, oxLDL/ß2GPI/aß2GPIAb complex induced an increase of ROS level during apoptosis, which could be attenuated by antioxidants. The effect of complex-induced apoptosis could also be reversed to some extent by antioxidants. Conclusion The oxLDL/ß2GPI/aß2GPIAb complex can induce the apoptosis in HUVECs by increasing ROS.

[oxLDL/ß2GPI/anti-ß2GPI antibody complex inhibits autophagy in RAW264.7 cells through the activation of AKT pathway].

Objective To investigate the effect of oxidative low-density lipoprotein/ß2 glycoprotein I/anti-ß2 glycoprotein I antibody (oxLDL/ß2GPI/anti-ß2GPI antibody) complex on autophagy in mouse macrophages (RAW264.7 cells) and its mechanism. Methods RAW264.7 cells were stimulated with medium, oxLDL, oxLDL/ß2GPI complex, oxLDL/ß2GPI/anti-ß2GPI antibody complex, oxLDL/anti-ß2GPI antibody complex and ß2GPI/anti-ß2GPI antibody complex separately. The protein levels of autophagy-associated proteins including LC3 and P62 were detected by Western blot analysis. The tandem mRFP-GFP-LC3 adernovirus was used to detect the patency of autophagosomes and autophagy flow. The AKT pathway inhibitor LY294002 (50 µmol/L) was used for investigating the role of AKT pathway in the autophagy in RAW264.7 cells treated by oxLDL/ß2GPI/anti-ß2GPI antibody complex. Results oxLDL/ß2GPI/anti-ß2GPI antibody complex reduced the level of LC3-II protein and the number of autophagosomes, increased the level of P62 protein and blocked the autophagy flow. In addition, the autophagy in RAW264.7 cells was partially restored by the pre-treatment of LY294002. Conclusion The oxLDL/ß2GPI/anti-ß2GPI antibody complex can inhibit the autophagy in RAW264.7 cells through activating the AKT pathway.

[OxLDL/ß2GPI/anti-ß2GPI complex promotes calcification and inflammatory cytokine expression in A7r5 cells].

Objective To evaluate the effect of oxidized low-density lipoprotein/ß2-glycoprotein I/anti-ß2 glycoprotein I antibody (oxLDL/ß2GPI/anti-ß2GPI) complex on the calcification and inflammatory cytokine expression in A7r5 rat vascular smooth muscle cells, and to find out the role of Toll-like receptor 4 (TLR4) signal in this process. Methods The A7r5 cells were intervened with oxLDL, oxLDL/ß2GPI complex, oxLDL/anti-ß2GPI complex, ß2GPI/anti-ß2GPI complex, and oxLDL/ß2GPI/anti-ß2GPI complex for different time, with or without TLR4 inhibitor (TAK-242). Alizarin red staining was used to observe the calcification. Real-time quantitative PCR was applied to detect the total mRNA levels of actin-associated protein SM22α (trangelin) and Runt-related transcription factor 2 (RUNX2). The protein levels of SM22α and RUNX2 were measured by Western blotting. Tumor necrosis factor α (TNF-α) was detected by ELISA. Different concentrations of TNF-α were adopted to stimulate A7r5 cells, and the above methods were operated to examine the changes of SM22 and RUNX2. Results When A7r5 cells were incubated by oxLDL/ß2GPI/anti-ß2GPI complex, more calcified nodules were observed, the expression of SM22α was reduced and RUNX2 expression was enhanced at both mRNA and protein levels. And the complex increased the expression of inflammation cytokine TNF-α. TLR4 inhibitor TAK-242 reversed the phenomenon. Different concentrations of TNF-α could reduce mRNA and protein expression of SM22α while raise RUNX2 expression. Conclusion oxLDL/ß2GPI/anti-ß2GPI complex can increase the expression of cytokine TNF-α, and thus quicken calcification procedure in A7r5 cells, along with the change of the traditional contraction phenotype into the osteogenic phenotype. TLR4 receptor participates in this process.

Association between Maternal Weight Indicators and Iron Deficiency Anemia during Pregnancy: A Cohort Study.

BACKGROUND: The effect of maternal weights on the risk of iron deficiency anemia (IDA) during pregnancy remains unclear. The study aimed to investigate the association between maternal weight indicators and IDA during pregnancy. METHODS: We conducted a cohort study to examine the association between maternal weight indicators, including prepregnancy body mass index and the rate of gestational weight gain (GWG), and the risk of IDA among Chinese pregnant women. Data about new-onset IDA at different trimesters from a national cross-sectional survey were collected; information regarding baseline variables and rate of GWG from women participating in the survey were retrospectively collected. Tested IDA and reported IDA were documented. Multilevel logistic regression to examine the association between maternal weight indicators and the risk of IDA after adjusting for potential confounders was conducted. RESULTS: This study enrolled 11,782 pregnant women from 24 hospitals from September 19, 2016, to November 20, 2016. Among those, 1515 (12.9%) IDA events were diagnosed through test (test IDA); 3915 (33.3%) were identified through test and patient reporting (composite IDA). After adjusting for confounders and cluster effect of hospitals, underweight pregnant women, compared with normal women, were associated with higher risk of test IDA (adjusted odds ratio [aOR]: 1.35, 95% confidence interval [CI]: 1.17-1.57 and composite IDA (aOR: 1.35, 95% CI: 1.21-1.51); on the contrary, overweight and obese women had lower risk of test IDA (aOR: 0.68, 95% CI: 0.54-0.86 overweight; aOR: 0.30, 95% CI: 0.13-0.69 obese) and composite IDA (aOR: 0.77, 95% CI: 0.67-0.90 overweight; aOR: 0.34, 95% CI: 0.21-0.55 obese). The higher rate of GWG was associated with higher risk of IDA (test aOR: 1.86 95% CI: 1.26-2.76; composite aOR: 1.54, 95% CI: 1.16-2.03). CONCLUSIONS: Pregnant women who are underweight before pregnancy and who have faster GWG are more likely to develop IDA. Enforced weight control during pregnancy and use of iron supplements, particularly among underweight women, may be warranted.

Hepatitis B surface antigen positivity during pregnancy and risk of gestational diabetes mellitus: A systematic review and meta-analysis.

Chronic hepatitis B virus (HBV) infection is a prevalent public health issue worldwide. Its impact on important pregnancy outcomes, such as gestational diabetes mellitus (GDM), has not been clearly established. The findings from published studies are inconsistent. In this systematic review and meta-analysis, we aimed to clarify whether HBV infection manifested during pregnancy is associated with an increased risk of GDM. We searched MEDLINE and EMBASE for cohort studies and case-control studies that investigated the association between maternal hepatitis B surface antigen (HBsAg) positivity and GDM. We pooled adjusted odds ratio (aOR) and unadjusted OR, respectively, using the random-effect generic inverse variance method. We assessed risk of bias using the Quality in Prognosis Studies tool and conducted five pre-specified subgroup analyses. In total, 23 cohort studies involving 3 529 223 participants were included. The risk of GDM was 6.48% (1868/28 829) among HBsAg-positive pregnant women and 3.41% (119 283/3 500 394) among HBsAg-negative pregnant women. Meta-analyses of both unadjusted and adjusted effect estimates showed that HBsAg positivity during pregnancy was associated with higher risk of developing GDM (unadjusted OR 1.35, 95% CI: 1.17 to 1.56, I2  = 82.6%; adjusted OR 1.47, 1.22 to 1.76, I2  = 62%). Among pre-specified subgroup analysis, significant differences were found among studies with high vs low or moderate risk of bias. The results were robust to sensitivity analyses. In conclusion, HBsAg positivity during pregnancy has a moderate effect on an increased risk of GDM. Given the size of the population with HBV infection worldwide, however, this effect could have substantial impact on pregnancy. Further studies are warranted to investigate whether active infection with HBeAg positivity would further elevate the risk of adverse events during pregnancy.

Effects of Toll-like receptor 4 on ß2-glycoprotein I-induced splenic T cell subsets differentiation.

Our previous study demonstrated that beta 2-glycoprotein I (ß2GPI) stimulation promotes bone marrow derived dendritic cells (BMDCs) maturation and T cell proliferation in a Toll-like receptor 4 (TLR4) dependent manner. However, ß2GPI induced T cell differentiation and the role of TLR4 in this process have rarely been reported. In the present study, we focused on the differentiation of splenic T cells in ß2GPI immunized Balb/c, C3H/HeN and C3H/HeJ mice. According to our results, Th2 dominated differentiation was observed in ß2GPI immunized Balb/c and C3H/HeN mice than in those treated with normal saline (NS), namely the up-regulated levels of Th2 markers GATA3 and IL-4 (p < 0.05). Meanwhile, reduced Th1 markers T-bet and IFN-γ, and Treg marker Foxp3 were observed in ß2GPI immunized mice (p < 0.05). C3H/HeJ mice have the same gene background with C3H/HeN mice except a functional mutant in TLR4 gene. However, the described Th2 differentiation was not detected in these TLR4 deficient mice, indicating the importance of TLR4 in immune response against ß2GPI. In addition, we found that ß2GPI-induced Th2 differentiation could be strengthened by cytokines secreted by dendritic cells (DCs) and DCs-T cells interaction. However, DCs-T cells contact was indispensable during this process because of its unique role in suppressing Th1 function. Furthermore, this Th2 biased differentiation pattern was more noticeable in mice received 4 times ß2GPI immunization than those received 2 times, suggesting the amplifying effects of anti-ß2GPI Ab on ß2GPI induced Th2 response. These findings may partly explain the immune imbalance in APS patient through the view angle of T cell differentiation and anti-ß2GPI antibody production.

[The ß2GPI/aß2GPI inhibits phagocytosis of oxidized low-density lipoprotein in THP-1 macrophages via activating TLR4].

Objective To investigate the effects of ß2 glycoprotein I/anti-ß2 glycoprotein I antibody complex (ß2GPI/aß2GPI) on lipid phagocytosis and class B scavenger receptor CD36 expression of macrophages, and the role of Toll-like receptor 4 (TLR4) during the process. Methods THP-1 macrophages were induced from THP-1 cells treated by 100 ng/mL phorbol ester (PMA), and then identified by morphological observation and DiI-oxLDL uptake assay. THP-1 macrophages were treated with RPMI1640 medium, ß2GPI, aß2GPI or ß2GPI/aß2GPI. The protein and mRNA expression of TLR4 were detected by Western blot analysis and real-time fluorescent quantitative PCR (qRT-PCR), respectively. After that, THP-1 macrophages were treated with RPMI1640 medium, oxidized low-density lipoprotein (oxLDL), oxLDL combined with ß2GPI/aß2GPI or oxLDL combined with LPS. Intracellular lipid deposition was observed by oil red O staining; the mRNA and protein expression of CD36 were detected by qRT-PCR, immunofluorescence and Western blot analysis. To unveil the role of TLR4 in this process, THP-1 macrophages were pre-treated with or without TLR4 inhibitor TAK-242 (1 µg/mL). Results After PMA treatment, THP-1 cells showed macrophage-like morphology and were able to engulf DiI-oxLDL. Compared with RPMI1640 medium, ß2GPI/aß2GPI treatment significantly increased TLR4 expression in THP-1 macrophages. Compared with oxLDL alone, oxLDL combined with ß2GPI/aß2GPI inhibited lipid deposition and CD36 expression in THP-1 macrophages, which could be partly reversed by TLR4 blockage. Conclusion The ß2GPI/aß2GPI can inhibit the phagocytosis of oxLDL and CD36 expression in macrophages, which is linked to the function of TLR4.

[The mechanism of oxLDL/ß2GPI/anti-ß2GPI antibody complex promoting the expression of adhesion molecules in HUVECs].

Objective To investigate the effect of oxidative low-density lipoprotein/ß2 glycoprotein I/anti-ß2 glycoprotein I antibody (oxLDL/ß2GPI/anti-ß2GPI-Ab) complex on the expressions of adhesion molecules in human umbilical vein endothelial cells (HUVECs), and the role of Toll-like receptor 4 (TLR4) signaling pathway during this process. Methods HUVECs were stimulated with simple medium, oxLDL, oxLDL/ß2GPI complex, oxLDL/anti-ß2GPI-Ab complex, oxLDL/ß2GPI/anti-ß2GPI-Ab complex and LPS separately, and then the total mRNA and protein were collected. The mRNA and protein expressions of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and von Willebrand factor (vWF) were detected by real-time quantitative PCR and Western blotting, respectively. Pre-treatment method was applied to explore the influence of TLR4 antagonist TAK-242 (5 µmol/L) and p38MAPK antagonist SB203580 (10 µmol/L) on the expressions of adhesion molecules in HUVECs. THP-1 cell adhesion test was employed to evaluate the effect of oxLDL/ß2GPI/anti-ß2GPI-Ab complex on the ability of attracting monocytes of HUVECs. Results Compared with simple medium, oxLDL/ß2GPI/anti-ß2GPI-Ab complex obviously upregulated the expressions of ICAM-1, VCAM-1 and vWF in HUVECs and promoted the adhesion of THP-1 cells to HUVECs. In addition, these effects were inhibited by the pre-treatment of TAK-242 or SB203580. Conclusion OxLDL/ß2GPI/anti-ß2GPI-Ab complex could up-regulate the expressions of adhesion molecules in HUVECs and promote the adhesion of THP-1 cells to HUVECs, in which TLR4 and p38 MAPK were closely involved.

[The oxLDL/ß2GPI/ß2GPI-Ab complex promotes the migration and the expression of inflammatory cytokines in human umbilical vein endothelial cells].

Objective To study the effects of the complex of oxidized low density lipoprotein/ß2-glycoprotein I/ß2-glycoprotein I antibodies (oxLDL/ß2GPI/ß2GPI-Ab) on the migration of human umbilical vein endothelial cells (HUVECs) and the expression of inflammatory cytokines, and their underlying Toll-like receptor (TLR4) pathway. Methods HUVECs were treated with oxLDL, oxLDL/ß2GPI complex, oxLDL/ß2GPI-Ab complex, oxLDL/ß2GPI/ß2GPI-Ab complex, or lipopolysaccharide (LPS) for a period of time in their corresponding groups. The migration of HUVECs was observed by the wound-healing assay. The mRNA and protein levels of TLR4 in HUVECs were detected by real-time quantitative PCR (qRT-PCR) and Western blotting, respectively. The cells were pretreated with or without TAK-242 (the inhibitor of TLR4) 2 hours before stimulated by corresponding stimulus as described above. Then, the contents of monocyte chemotactic protein-1 (MCP-1), interleukin 1ß (IL-1ß) and IL-6 in cell culture supernatant were determined by ELISA, and their mRNAs were detected by qRT-PCR. Results The oxLDL/ß2GPI/ß2GPI-Ab complex promoted the migration of HUVECs effectively, and increased the expression of TLR4. The oxLDL/ß2GPI/ß2GPI-Ab complex increased the expressions of MCP-1, IL-1ß, and IL-6. TAK-242 could reduce the effects of oxLDL/ß2GPI/ß2GPI-Ab complex. Conclusion The oxLDL/ß2GPI/anti-ß2GPI-Ab complex can promote the migration of HUVECs and the expression of related inflammatory cytokines, and TLR4 may be involved in this process.

[Anti-ß2GPI antibody promotes release of inflammatory and pro-thrombosis molecules from arteries in apolipoprotein E-deficient mice].

Objective To investigate the roles of anti-beta 2 glycoprotein I antibodies (anti-ß2GPI Ab) in the expressions of atherosclerosis(AS)-related inflammatory factors and pro-thrombosis molecules in apolipoprotein E-deficient (ApoE-/-) mice. Methods ApoE-/- mice were randomly divided into normal saline (NS) group, 100 µg anti-ß2GPI Ab group, 100 µg homologous antibody (rabbit-IgG) group and 100 µg ß2GPI/anti-ß2GPI Ab complex group after silastic collars were placed around their carotid arteries by surgery. All mice were fed a high fat diet and corresponding stimuli were given through intraperitoneal injection at 7-day intervals. Six weeks later, the mice were executed. The blockage of carotid arteries of the operated side was observed by HE staining. The expressions of TLR4, tissue factor (TF) and von Willebrand factor (vWF) were detected by immunohistochemistry. The mRNA levels of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in aorta were tested by real-time quantitative PCR. Results HE staining showed that the blockage of carotid arteries in antibody group was the most obvious. The immunohistochemistry showed that the expressions of TLR4, TF and vWF in anti-ß2GPI Ab group increased remarkably. Furthermore, the mRNA levels of IL-1ß and TNF-α in anti-ß2GPI Ab group were higher than those in the other groups. Conclusion The anti-ß2GPI antibody promotes the formation of atherosclerotic plaques in mice by up-regulating the release of inflammatory cytokines IL-1ß, TNF-α and thrombosis-related molecules TF, vWF and TLR4, ultimately enhancing the development of AS.